Modulators of cholecystokinin

ABSTRACT

CCK modulators, e.g. agonists or antagonists, of the following formula (I): ##STR1## or a base-addtion salt thereof.

This is a continuation of U.S. Ser. No. 07/914,918, filed Jul. 14, 1992,now U.S. Pat. No. 5,380,872.

FIELD OF THE INVENTION

The invention relates to chemical compounds which modulate the hormonecholecystokinin (CCK) in mammals. Effects of such modulation include theregulation of appetite, and treatment of gastrointestinal disorders(including gallbladder disorders), central nervous system disorders andpain.

BACKGROUND OF THE INVENTION

CCK is a gastrointestinal hormone of 33 amino acids which is utilized bythe body in the cascade of events which are part of hunger, eating,digestion and satiety. Other such hormones include gastrin and secretin.The presence of acid, fat and protein breakdown products or anyirritating factor in the upper small intestine causes the release ofsecretin and CCK. Both of these are important for control of pancreaticsecretion and CCK is essential for emptying of the gallbladder, seeChapter 64, entitled "Secretory Functions of the Alimentary Tract" in A.C. Guyton's Textbook of Medical Physiology, W. B. Saunders, 1986Philadelphia.

CCK has a variety of regulatory roles in the periphery includinggallbladder contraction and pancreatic enzyme secretion (V. Mutt in"Gastrointestinal Hormones", G. B. J. Glass, ed, Raven Press, New York,1980 pp. 169; J. A. Williams, Biomed. Res., 1982 pp. 3:107), inhibitionof gastric emptying and suppression of food intake. CCK and itsfragments are believed to play an important role in appetite regulationand satiety (Della-Fera, Science, 1979 pp. 206:471; Saito et al.,Nature, 1981 pp. 289:599; and Smith "Eating and its Disorders", A. J.Stunkard and E. Stellar, eds., Raven Press, New York, 1984 p. 67) andrecently, patients with bulimia were shown to have lower than normal CCKlevels in their plasma (Geracioti et al., New England Journal ofMedicine, 1988 pp. 319:683).

CCK in the brain has been suggested to have a role in schizophrenia (N.P. V. Nair et al, Prog. Brain Res, 1986 pp. 65:237), memory andcognition (S. Itoh and H. Lal, Drug Dev. Res., 1990, pp. 21:257), andCCK antagonists have been suggested to be potentially useful in drugabuse therapy (B. Costall et al. in "Proceedings of the CambridgeSymposia, The Neurological Basis of Anxiety," Robinson College,Cambridge, U.K., Sep. 7 and 8, 1990).

Two sub-types of the CCK receptor have been identified. Type-A CCKreceptors, commonly referred to as the "peripheral-type" receptor, areprimarily found in the pancreas, gallbladder, ileum, pyloric sphincterand on vagal afferent nerve fibers. Type-A CCK receptors bind CCK-8 withhigh affinity but have low affinity for desulfated CCK-8 and CCK-4. Thebrain contains predominantly the Type-B receptors that bind CCK-8,desulfated CCK-8 and CCK-4 with high affinity. Type-A CCK receptors arefound in the brain, although in low abundance (D. R. Hill et al., BrainRes, 1988, pp. 454:101-5; D. R. Hill et al., Neurosci Lett., 1988, pp89:133-9; R. W. Barrett et al., Mol. Pharmacol, 1989, 36:285-90; and D.R., Hill et al., J. Neurosci, 1990, 10:1070, 81), and play an importantrole there also (V. Dauge et al., Pharmacol Biochem Behav., 198933:637-40). Type-A receptor-selective CCK agonists are currently ofparticular interest as potential anorectic agents because of the abilityof CCK-8 and Type-A CCK-selective agonists to suppress food intake inseveral animal species (Della-Fera et al., Science, 1979, 206:471; K. E.Asin et al., Intl Conference on Obesity., 1990, Abstract p.40).

Obesity is a major disorder affecting as much as one third of the NorthAmerican population. Several studies have shown that such individualsare at increased risk in developing cardiovascular disease (hypertensionand hypercholesterolemia), diabetes and several types of cancer. Theeffective treatment of obesity, however, remains a largely unachievedgoal. Existing pharmacotherapeutic approaches to weight loss involve theuse of amphetamine-based agents such as amphetamine, diethylpropion,mazindol and fenfluramine which act directly on the CNS to lower foodintake by modulating dopaminergic, adrenergic and/or serotonergicmechanisms. Although weight loss can be achieved with such agents, theiruse is restricted due to CNS side-effects, potential addiction liabilityand the production of tolerance to their actions, with chronicadministration leading to potential depression, vestibular disturbances,hallucinations and addiction, as well as interference with the actionsother drugs such as MAO inhibitors and antihypertensives. There is alsoa subpopulation of obese patients that is refractory to presentanorectic drug treatments.

Additional roles for CCK in the periphery (body) include stimulation ofgall bladder contraction, and inhibition of gastric emptying. CCK in thebrain has been suggested to have a role in schizophrenia, memory andcognition and CCK antagonists have been suggested to be useful in drugabuse therapy, all as set forth in the previous citations or incitations therein.

CCK agonists or analogs of CCK-8, a particular biologically active formof CCK, have been published. For example, U.S. Pat. No. 4,490,364,issued 25 Dec. 1984 discloses heptapeptide, octapeptide and nonapeptideanalogs of CCK-8 as CCK agonists for stimulating gallbladdercontractions, arresting the secretion of gastric acid and treatingconvulsions.

European Patent Application EP381,340, published 8 Aug. 1990, andEuropean Patent Application EP268,297, published 25 May 1988, disclosehepta- and octapeptides with sulfate ester groups which are useful fortreating obesity.

Orally active CCK-B receptor selective dipeptoid antagonists aredescribed in European Patent 465,537 published 2 Jan. 1991 and by D. C.Horwell et al., in J. Med. Chem., 34, 1991, pp. 404-414. These compoundshave weak CCK-A receptor affinity, being >500-fold selective for CCK-Breceptors, and are reported to be useful as anti-anxiety and anti-ulceragents. These compounds also possess weak anorectic activity resultingfrom their weak affinity for CCK-A receptors. Six related disclosureshave been published. PCT WO 92/04348 published Mar. 19, 1992 teachescarboline derivatives useful in treating obesity while cholecystokininantagonists are described in PCT WO 92/04045 and WO 92/04025. Dipeptoidsto treat obesity are taught in PCT WO 92/04322 while pro-drugs todipeptoids are shown in PCT WO 92/04038. Peptoids for treating obesityare found in PCT WO 92/04320.

CCK antagonists or gastrin receptor antagonists comprising C-terminalfragments of CCK have recently been reported. See Jensen et al.,Biochem. Biophys. Acta, 1983, pp. 757-250; Spanarkel, J. Biol. Chem.1983, pp. 258-6746. Japanese Patent Application 45/1050 to Miyao et al.discloses a tetrapeptide derivative of the carboxy terminal sequence ofgastrin (L-Trp-L-Lys-L-Asp-L-PheNH₂) which acts as antagonists ofgastrin.

CCK-A receptor selective tetrapeptide agonists are described in theliterature as agents to treat gastrointestinal disorders (including gallbladder stasis), CNS disorders and pain as well as appetite regulation,see PCT WO 91/19733 published 26 Dec. 1991, by K. Shiosaki et al., in J.Med. Chem., 33, 1990 pp. 2950-2952 and in PCT WO 90/06937, published 28Jun. 1990.

The present invention describes CCK-A receptor selective agents whichmodulate peripheral CCK receptors.

SUMMARY OF THE INVENTION

Cholecystokinin receptor ligands of the formula (I): ##STR2## or apharmaceutically acceptable salt thereof, wherein A, B, D, E and G andm, q, and R¹², R¹⁵, R¹⁶, and R¹⁸ a are defined herein, are useful asmodulators of CCK with specific utilities including the induction ofweight loss. Also claimed are pharmaceutical compositions of thesecompounds and methods for their use as pharmaceuticals, intermediates intheir synthesis as well as the synthesis itself,

DETAILED DESCRIPTION OF THE INVENTION

Compounds of formula (I) above are part of the invention where:

A is selected from

--COOR¹,

--CONHR¹,

--CN,

--CHN₄, i.e. C-linked tetrazole,

--CONHSO₂ R²,

--CH₂ OR¹,

--CH₂ SR¹,

--P--O(OR¹)₂,

--CO--NHOH, ##STR3##

R¹ is H or lower alkyl;

R² is lower alkyl, fluoro lower alkyl, aryl, substituted aryl or OH;

R³ is OH, NH₂ or CH₃ ;

R⁴ is CN, CO₂ H or CF₃ ;

B is selected from

i) --COCH(NHR⁵)--(CH₂)_(n) --;

ii) --COCH₂ (CH₂)_(n) --;

iii) --CO--CH═CH--, cis or trans;

iv) --CO--phenylene--, ortho, meta or para;

v) --CH₂ --(CH₂)_(n) --;

vi) --CH(COOR⁶)--CH₂ --(CH₂)_(n) --;

where n is 0, 1 or 2;

R⁵ is H, lower alkyl, COR⁷ or CONHR⁸ ;

R⁶ is H, lower alkyl, aryl, substituted aryl or arylalkyl;

R⁷ is H, lower alkyl, aryl, substituted aryl or arylalkyl;

R⁸ is H, lower alkyl, aryl, substituted aryl or arylalkyl;

R⁹ is H or lower alkyl;

R¹⁰ is H or lower alkyl;

m is 1,2, 3 or 4;

G is --O--, --CH₂ --, --NR¹¹ --, --CH═CH--, cis or trans;

R¹¹ is H or lower alkyl;

R¹² is selected from

i) alkyl;

ii) alkenyl;

iii) cyclo (C₃ -C₁₀ alkyl);

iv) heterocyclic;

v) substituted heterocyclic;

vi) arylalkyl;

v) aryl, having 1 or 2 substituents independently selected from thegroup consisting of:

a) hydroxyl,

b) halogen,

c) --OSO₃ ^(R) ¹³,

d) nitro,

e) cyano,

f) amino,

g) lower alkylamino,

h) (lower alkyl)2 amino,

i) lower alkyl,

j) halo lower alkyl,

k) lower alkoxy,

l) C₂ -C₄ -alkanoyl,

m) lower alkoxy carbonyl, and

n) phenoxy

R¹³ is H or lower alkyl;

p is 0, 1,2;

D is

i) --NH--CO--,

ii) --CO--NR¹⁴ --,

iii) --(CH₂)_(r) --NR¹⁴ --,

v) --NR¹⁴ --(CH₂)_(r) --

r is 0, 1 or 2

R¹⁴ is H or lower alkyl;

R¹⁵ is H or lower alkyl;

q is 0 or1;

R¹⁶ is selected from

i) aryl or substituted aryl,

ii) substituted aryl,

iii) heteroaryl,

iv) substituted heteroaryl, or

v) bicyclic heteroaryl;

E is selected from

i) --NH--CO--,

ii) --CO--NR¹⁷ --,

iii) --NH--CO--NR¹⁷ --,

iv) --O--CO--NR¹⁷ --,

v) --SO₂ --NR¹⁷ --, or

vi) --(CH₂)_(r) --NR¹⁷ --;

R¹⁷ is H or lower alkyl,

R¹⁸ is selected from

i) C₁ -C₁₀ alkyl or C₂ -C₁₀ alkenyl,

ii) C₃ -C₁₀ mono, bi- or tri-cycloalkyl with zero to four substituentsindependently selected from the group consisting of

a) alkyl,

b) halogen,

c) CN,

d) --OR¹⁹,

e) --SR¹⁹,

f) --CO₂ R¹⁹,

g) --CF₃,

h) --NR¹⁹ R²⁰,

i) --(CH₂)_(s) OR¹⁹, and

j) --(CH₂)_(s) COOR¹⁹ ;

iii) heterocyclic,

iv) substituted heteroaryl,

v) arylalkyl,

vi) aryl,

vii) aryl having 1 or 2 substituents independently selected from s is aninteger from 0 to 6;

R¹⁹ is H or lower alkyl; and

R²⁰ is H or lower alkyl.

The pharmaceutically acceptable salts of the acids of formula (I) arereadily prepared by conventional procedures such as treating an acid offormula (I) with an appropriate amount of a base, such as an alkali oralkaline earth metal hydroxide e.g. sodium, potassium, lithium, calcium,or magnesium, or an organic base such as an amine, e.g.,dibenzylethylenediamine, cyclohexylamine, dicyclohexylamine,trimethylamine, piperidine, pyrrolidine, benzylamine and the like, or aquaternary ammonium hydroxide such as tetramethylammonium hydroxide andthe like.

In the definitions for the above formula (I), lower alkyl may be of 1-6carbons, straight or branched chain; alkyl may be of about 1-10 carbons,straight or branched chain such as but not limited to methyl, ethyl,n-propyl, iso-butyl or 3-methyl pentyl; alkenyl may be from about 2 to 6carbons; halogen may be fluoro, chloro, bromo or iodo; the definitionsfor divalent moieties such as B, D, E and G are read exactly as shownabove into formula (I) e.g. if B is --COCH₂ (CH₂)--, the --CO-- portionis attached to the NH of (I) while the --(CH₂)-- is attached to the Amoiety.

Particular compounds of Formula (I) are those of the following formulae(Ia), (Ib), (Ic), (Id) and (Ie), and wherein * is R or S and ** is R orS and preferably * is R and ** is R or S: ##STR4## where R¹⁸ is benzyl,adamantyl, t-butyl or trans-2-methylcyclohexyl; and ##STR5## where R¹⁵is methyl and R¹⁶ is 3-quinolyl, 2-naphthyl, 3-indolyl, 2-indolyl orphenyl; or R¹⁵ is hydrogen and R¹⁶ is 3-indazolyl, 3-quinolyl, 2-naphyl,3-indolyl, 2-indolyl or phenyl; and ##STR6## where G is --O-- or --NH--and R¹² is phenyl, 2-methylphenyl, 2-chlorophenyl, 2-methoxyphenyl or2-nitrophenyl; or G is --CH═CH-- and R¹² is phenyl, 4-methylphenyl,4-chlorophenyl, 4-methoxyphenyl, 4-hydroxyphenyl or 4-nitrophenyl; or Gis --CH₂ -- or --CH₂ CH₂ -- and R¹² is any of the values given above forR¹² Formula (I); and ##STR7## wherein B is (CH₂)_(n), n=1, 2, 3, CH═CH,cis or trans and A is CO₂ H, CO₂ CH₃, CHN₄, NHSO₂ CF₃ or NHSO₂ CH₃ ; and##STR8##

Preferred for agonist activity is R¹⁵ =CH₃ ; m=3, 4; and ** is R or S.

Preferred for antagonist activity is R¹⁵ =H; m=1-4; * is R; ** is R orS.

Compounds of formula (I) are CCK-A modulators, e.g. antagonists oragonists, and would be useful in the treatment and prevention ofCCK-related disorders of the gastrointestinal and appetite systems ofanimals, especially man. CCK-A agonists would be useful in the treatmentof obesity or gall bladder disorders e.g. gall stones, while CCK-Aantagonists would be useful in the treatment of pancreatic cancer, andas central nervous system suppressants, e.g. anti-psychoties.

The present invention is also directed to pharmaceutical compositionscomprising a therapeutically--effective amount of the compound offormula (I) and a pharmaceutically--acceptable carrier or diluent, aswell as to a method of treating gastrointestinal disorders (includinggallbladder disorders), CNS disorders, insulin-related disorders andpain, or of regulating appetite in humans and lower mammals, byadministration of a compound of formula (I).

Preferred embodiments of this invention are when R¹⁸ is a substituted orunsubstituted C₆ to C₁₀ cycloalkyl or polycycycloalkyl of the followingformulae (i)-(iv): ##STR9## wherein W, X, Y, and Z are independently H,C₁ -C₆ straight or branched alkyl, CF₃,NR¹⁹ R²⁰,(--CH₂)_(s) COOR¹⁹, CN,F, Br, CI, OR¹⁹ or SR¹⁹.

Preferred compounds for CCK-A antagonists are R¹⁵ =H, m=1,2, 3, 4.

Preferred compounds for CCK-A agonist activity are R¹⁵ =CH₃, m=3, 4.

Also preferred is when R¹⁶ is indole, napthyl or quinoyl. Also preferredis when p=1, E═OCONH and D═CONH.

Particular subgroups of formula (I) are those wherein:

i) --B--A is --CO--CH₂ CH₂ COOH; --COCH₂ (CH₂)_(n) --NHSO₂ CF₃ where nis 0 or 1; or

ii) --G--R¹² is --NH phenyl with CI, OCH₃ or CH₃ substitution at theortho position; and/or

iii) R¹⁸ is adamantane, norbornane, 2-methylcyclohexyl or tert-butyl.

The compounds of the present invention can have multiple chiral centers,depending on their structures, and may exist as diastereomers, mixturesof diastereomers, or as a mixture of the individual enantiomers. Thepresent invention contemplates all such forms of the compounds. Themixtures of is diastereomers are typically obtained as a result of thereactions described more fully below. Individual diastereomers may beseparated from mixtures by conventional techniques such as columnchromatography or repetitive recrystallizations. Individual enantiomersmay be separated by conventional methods such as conversion to a saltwith an optically active compound followed by recrystallization.Alternatively, chiral chromatography may be used.

Individual a-amino acids are known, or if not known, may be synthesizedand resolved by methods within the skill of the art.

Two key intermediates are essential to the present invention: triaminointermediate (II) and b-diamino acid (III): ##STR10##

The intermediates (II) and (III) are derived from naturally occurring L-or D- a-amino acids (m=3,4) or synthetic a-amino acids (m=1,2). Wherem=4, N-a-protected L or D lysine may be purchased. Where m=3,N-a-protected L- or D- ornithine may be purchased. Where m=1, 2,compounds are obtained by the following route illustrated in Scheme 1for L- or D- asparagine, or L- or D- glutamine, respectively, using themethod of Waki, et al (Synthesis (1981) 266-268). ##STR11##

Wherein BOC is t-butyloxycarbonyl.

Scheme I above describes the synthesis of differentially protected aminoacids of formula VI, which involves treatingN-a-t-butyloxycarbonyl-asparagine or glutamine withiodobenzenetrifluoroacetate in aqueous N,N-dimethylformamide to effectthe Hofmann reaction. The free amine, compound V, is then suitablyprotected with benzyl chloroformate and triethylamine to give thecarbobenzyloxycarbamate. N-a-t-butyloxycarbonyl-ornithine or lysine canbe similarly protected, at the d-NH or e-NH, respectively.

The synthesis of a suitably protected triamino intermediate (II) isillustrated in Scheme 2: ##STR12##

Scheme 2 above illustrates procedures for preparing intermediates usefulin producing final products of Scheme 6.

Key intermediate Ila is prepared by first reducing the carboxylic acidof compound VI to an alcohol. In one process the acid is activated withisobutylchloroformate (iBuCF) and Hunig's base in a solvent such as THFat about 0° C. In another process the acid is treated with carbonyldiimidazole (CDI) in THF at room temperature. The activated ester orimidazolide is then reduced to an alcohol with a reducing reagent suchas sodium borohydride or lithium aluminum hydride. The lo alcohol,dissolved in dichloromethane, is treated with methanesulfonyl chlorideand triethylamine to give the corresponding mesylate. The mesylate istreated with sodium azide in N,N-dimethylformamide (DMF) to produce theazide. The azide is hydrogenated over a Lindlar catalyst to form amineIIa.

Alternatively, the side chain nitrogen may be functionalized prior tothe synthesis of the triamino intermediate. This synthesis isillustrated in Scheme 3, where J=NH, CH₂, particular values of G,N-a-t-butyloxycarbonyl (as illustrated), or the N-a-benzyloxycarbonylprotecting groups may be used. ##STR13##

Scheme 3 above illustrates procedures for preparing intermediates usefulin producing final products.

One process involves treating N-a-t-butyloxycarbonyl-lysine withisocyanates in 1N sodium hydroxide at room temperature. The carboxylicacid is activated with such reagents as isobutylchloroformate (iBuCF)and Hunig's base or carbonyl diimidazole (CDI) in THF and the ester orimidazolide is reduced to an alcohol with reagents such as sodiumborohydride or lithium aluminum hydride.

The alcohol, dissolved in dichloromethane, is treated withmethanesulfonyl chloride and triethylamine to give the correspondingmesylate. The mesylate is treated with sodium azide inN,N-dimethylformamide to produce the corresponding azide. The azide ishydrogenated in the presence of a catalyst such as ten percent palladiumon carbon to form amine (IIIb).

The synthesis of a suitable protected b-diamino acid intermediate (III)is illustrated in Scheme 4. ##STR14##

Scheme 4 above illustrates procedures for preparing intermediates usefulin producing final products of Scheme 7.

In one process differentially protectedN-a-t-butyloxycarbonyI-N-e-carbobenzyloxylysine is homologated using theArndt Eistert reaction. The acid is treated with isobutylchloroformate(iBuCF) and triethylamine in a solvent such as THF followed by additionof diazomethane to give the corresponding diazomethylketone. A methanolsolution of the diazomethylketone is treated with silver benzoate toobtain the homologated methyl ester. This then is treated with lithiumhydroxide and stirred at room temperature overnight to produce thecorresponding carboxylic acid.

Alternatively, the side chain may be functionalized prior to synthesisof intermediate (III), as illustrated in Scheme 5, where J=NH, CH₂,particular values of G. ##STR15##

Scheme 5 above illustrates procedures for preparing intermediates usefulin producing final products.

In one process N-a-t-butyloxycarbonyI-N-e-o-tolylaminocarbonyl-lysine ishomologated using the Arndt Eistert reaction. The acid is treated withisobutylchloroformate (iBuCF) and triethylamine in a solvent such as THFfollowed by addition of diazomethane to give the correspondingdiazomethylketone. A methanol solution of the diazomethylketone istreated with silver benzoate to obtain the homologated methyl ester.This then is treated with lithium hydroxide and stirred at roomtemperature overnight to produce the corresponding carboxylic acid.

Intermediates (IIa) and (IIIa) can be used to build the molecules,allowing flexible introduction of side chain substituents. Protected L-or D-amino acids can be coupled to intermediate IIa and L- or D- aminoacid amides can be coupled to intermediate IIIa, utilizing knownliterature methods. Suitable coupling agents include dicyclohexylcarbodiimide (DCC), 1-(3-dimethylaminopropyl)-3-ethyl carbodiimidehydrochloride (EDC), or bis(2-oxo-3-oxazolidinyl) phosphinic chloride.N-Hydroxybenzotriazole (HOBt) or N-hydroxy succinimide (NHS) may beadded to improve coupling rates and inhibit racemization. Suitablesolvents include dichloromethane (DCM) and dimethylformamide (DMF). Anexample of such coupling is shown in the following Schemes 6 and 7.##STR16##

Where E, R¹⁵, R¹⁶ and R¹⁸ are as previously defined.

In one process a solution of2-adamantyloxycarbonyl-α-methyl-D,L-tryptophan in dichioromethane reactswith an equimolar solution of 1-hydroxybenztriazole, key intermediate(IIa), and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride.The reaction mixture is allowed to react at room temperature overnightto form the carbobenzyloxy-protected product (XX). ##STR17##

Where E, R¹⁵, R¹⁶ and R¹⁸ are as previously defined.

In Scheme 7 a solution of2-adamantyloxycarbonyl-a-methyl-(D,L)-tryptophan in dichloromethanereacts with an equimolar solution of 1-hydroxybenztriazole, keyintermediate IIIa, and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimidehydrochloride. The reaction mixture is allowed to react at roomtemperature overnight to form the carbobenzyloxy protected product(XXII).

Intermediates (XX) and (XXII) may be deprotected using acid, such as HClor TFA, and the C-terminal group introduced either by reaction withanhydrides, amino acid active esters, or by coupling with appropriateamino acid esters, diacid half esters, or other suitable derivativesconsistent within the compounds of the invention as shown in thefollowing Scheme 8 wherein (XXIII) is an intermediate within Formula(XX) or (XXII): ##STR18##

In one procedure compound (XXIII) was treated with 4N hydrochloric acidin dioxane to obtain the hydrochloride salt of (XXIV). This then isdissolved in dichloromethane with triethylamine and reacted withsuccinic anhydride to form the succinyl free acid.

Intermediate (XXV) may be deprotected using H₂ and a suitable catalyst;or HBr/HOAc to give (XXVI) which may be heated with isocyanates, acidhalides, carbamoyl halides, cinnamoyl halides, alcohols with CDI, togive compounds of Formula (I) according to Scheme 9: ##STR19##

In another process compound (XXV) was hydrogenated in the presence of acatalyst such as ten percent palladium on carbon to form the amine XXVI.Alternatively, where the side chain functionality is defined,intermediates (IIb) and (IIIb) may be reacted in the same syntheticschemes to give a compound of Formula (XXVII): ##STR20## where K isoxycarbonylamino or oxycarbonyl and R²¹ is fluorenyl or benzyl, or C₁-C₂ alkyl.

After deprotection of the ester or amine, intermediates (XXVII) and(XXIX) can be used to prepare compounds of claim (I) through reaction of(XXVIII). ##STR21## with an appropriate amine in the presence of adehydrating agent such as DCC, EDC or BOP; or the reaction of formula(XXIX): ##STR22## with carbamoyl chlorides, alcohol with CDI, acidchlorides, acid anhydrides, protected amino acids, arthydrides,isocyanates.

When a compound of formula (I) is used as an agonist of CCK in a humansubject, the total daily dose administered in single or divided dosesmay be in amounts, for example, from 0.001 to 1000 mg a day and moreusually 1 to 1000 mg. Dosage unit compositions may contain such amountsof submultiples thereof to make up the daily dose.

The amount of active ingredient that may be combined with the carriermaterials to produce a single dosage form will vary depending upon thehost treated, the particular treatment and the particular mode ofadministration.

It will be understood, however, that the specific dose level for anyparticular patient will depend upon a variety of factors including theactivity of the specific compound employed, the age, body weight,general health, sex, diet, time of administration, rate of execretion,drug combination and the severity of the particular disease undergoingtherapy.

The compounds of the present invention may be administered sublingually,orally, parenterally, by inhalation spray, rectally or topically indosage unit formulations containing conventional nontoxicpharmaceutically acceptable carriers, adjuvants, and vehicles asdesired. Topical administration may also involve the use of transdermaladministration such as transdermal patches or iontophoresis devices. Theterm parenteral as used herein includes subcutaneous injections,intravenous, intramuscular, intrasternal injection, or infusiontechniques.

Injectable preparations, for example, sterile injectable aqueous oroleagenous suspensions may be formulated according to the known artusing suitable dispersing or wetting agents and suspending agents. Thesterile injectable preparation may also be a sterile injectable solutionor suspension in a nontoxic parenterally acceptable diluent or solvent,for example, as a solution in 1,3-butanediol. Among the acceptablevehicles and solvents that may be employed are water, Ringer's solution,and isotonic sodium chloride solution. In addition, sterile, fixed oilsare conventionally employed as a solvent or suspending medium. For thispurpose any bland fixed oil may be employed including synthetic mono- ordiglycerides. In addition, fatty acids such as oleic acid find use inthe preparation of injectables.

Suppositories for rectal administration of the drug can be prepared bymixing the drug with a suitable nonirritating excipient such as cocoabutter and polyethylene glycols which are solid at ordinary temperaturesbut liquid at the rectal temperature and will therefore melt in therectum and release the drug.

Solid dosage forms for oral administration may include capsules,tablets, pills, powders and granules. In such solid dosage forms, theactive compound may be admixed with at least one inert diluent such assucrose lactose or starch. Such dosage forms may also comprise, as innormal practice, additional substances other than inert diluents, e.g.,lubricating agents such as magnesium stearate. In the case of capsules,tablets and pills, the dosage forms may also comprise buffering agents.Tablets and pills can additionally be prepared with enteric coatings.

Liquid dosage forms for oral administration may include pharmaceuticallyacceptable emulsion, solutions, suspensions, syrups and elixirscontaining inert diluents commonly used in the art, such as water. Suchcompositions may also comprise adjuvants, such as wetting agents,emulsifying and suspending agents and sweetening, flavoring andperfuming agents.

The present agents can also be administered in the form of liposomes. Asis known in the art, liposomes are generally derived from phospholipidsor other lipid substances. Liposomes are formed by mono- ormulti-lamellar hydrated liquid crystals that are dispersed in an aqueousmedium. Any non-toxic, physiologically acceptable and metabolizablelipid capable of forming liposomes can be used. The present compostionsin liposome form can contain, in addition to the tetrapeptide of thepresent invention, stabilizers, preservatives, excipients and the like.The preferred lipids are the phospholipids and the phosphatidyl cholines(lecithins) both natural and synthetic.

Methods to form liposomes are known in the art. See, for example,Prescott, Ed., Methods in Cell Biology, Volume XIV, Acedemic Press, Newyork, N.Y. (1976), p. 33 et seq.

CCK-A RECEPTOR BINDING ASSAY

Tissue Preparation:

Solutions of 0.3M Sucrose and 2.0M Sucrose are prepared and chilledovernight at 4° C. On the following day inhibitors are added such thatthe final concentrations are 0.01% Soybean Trypsin Inhibitor (50mg/500mlsucrose) and 100 mM Phenylmethlysulfonyl fluoride (8.5 mg/500 mLsucrose).

Rats are sacrificed by decapitation using a guillotine. The rat'sexternal abdominal wall is wetted with methanol, and fur and skin areremoved. The abdomen is opened, the pancreas is carefully dissected out,and placed in a 50 mL beaker containing 0.3M sucrose. After all thepancreata have been harvested, excess fat and lymph nodes are trimmedoff. Pancreatic tissue is divided into approximately 4.0 g aliquots into30 mL beakers, each containing 1.0 mL of 0.3M sucrose.

In 4° C. cold room, the pancreata are minced with scissors and diluted1:10 weight:volume with 0.3M sucrose. Aliquots are homogenized in achilled 40 mL Wheaton dounce with 4 up and down strokes of the "B"pestle followed by 4 up and down strokes of the "A" pestle. Homogenatesare filtered through 2 layers of cheesecloth into a chilled 500 mLbeaker, then diluted with 2.0M sucrose with stirring to yield a finalconcentration of 1.3M sucrose homogenate. This 1.3M homogenate isdispensed into 18 thin-walled 36 mL polyallomer tubes on ice(approximately 30 mL homogenate per tube), and each tube is overlaidwith 0.3M sucrose until liquid is approximately 0.5 cm from the top ofthe tube. The samples are spun in a Sorvall RC70 ultracentrifuge at27,500 RPM (100,000×g) for 3 hours at 4° C. The interface band iscollected into a chilled graduated cylinder, diluted and mixed with colddistilled water to a total volume of 312 mL, and spun at 100,000×g for50 min at 4° C. The pellets are resuspended in KRH buffers, transferedto a 15 mL Wheaton dounce, and homogenized with 4 up and down strokes ofthe matched "A" (tight) pestle. This homogenate is transferred into 2-27mL polycarbonate bottles and spun at 100,000×g for 30 min at 4° C. Thepellet is resuspended (1 mL KRH buffer/gm wt of original tissue),transferred to an appropriate size dounce and homogenized with 4 up anddown strokes of the matched "A" pestle. 1 mL aliquots are stored at -70°C. in microcentrifuge tubes.

    ______________________________________                                        KRH Buffer: pH = 7.4 at 4° C.                                          COMPONENT        MW      g/1 L                                                ______________________________________                                        25 mM HEPES      260.3   6.51                                                 104 mM NaCl      58.44   6.08                                                 5 mM KCl         74.56   0.37                                                 1 mM KPO.sub.4   136.09  0.14                                                 1.2 mM MgSO.sub.4                                                                              246.48  0.30                                                 2 mM CaCl.sub.2  110.99  0.22                                                 2.5 mM Glucose   180.16  0.45                                                 0.2% BSA         --      2.00                                                 0.1 mM PMSF*     174.2   0.017                                                0.01% STI*       --      0.10                                                 ______________________________________                                         *inhibitors added fresh the day of the experiment                        

Assay:

Test compounds are diluted in assay binding buffer in stockconcentrations 10-fold more concentrated than desired final assayconcentration.

50 mL test compound+400 mL buffer+25 uL [¹²⁵ ]sulphate_(d) CCK-8labelled with Bolton and Hunter reagent (Amersham, 2000 Ci/mmol)+25 mLprepared rat pancreas membranes are incubated for 30 minutes at 25° C.while shaking gently throughout the incubation.

1 mM L-364718 (final concentration) is used for determination ofnon-specific binding.

Reaction is stopped using Brandell Cell Harvester, washing 3X with 3 mLice-cold (4° C.) assay binding buffer per wash.

Tissues are collected on Whatman GF/B filter papers pre-wet with assaybuffer and filter papers are counted using a gamma counter.

CCK-B RECEPTOR BINDING ASSAY

Tissue Preparation:

Hartley Male Guinea Pigs (250-300 g, Charles River) are sacrificed bydecapitation. The brain is removed and placed in 4° C. BUFFER (BUFFER=50mM Tris/HCL, pH=7.4). The cortex is dissected and placed in 4° C.BUFFER. The total wet weight of all cortices is determined and thetissues are diluted 1:10 (wt:vol) with BUFFER.

The cortex is minced using a Tekmar Tissuemizer, then homogenized inBUFFER with 5 up/down strokes using a motor driven glass/teflonhomogenizer. The preparation is maintained at 4° C. (on ice).

Membranes are pelleted by centrifugation in Sorvall RC5C at 4° C. usinga SA600 rotor spun at 16,000 RPM (47,800×g Maximum). The pellet is savedand the supernatent is discarded. The pellets are combined andresuspended in buffer at 4° C. using same volume as above and blended,as above with 5 up/down strokes of a glass/teflon motor drivenhomogenizer using the same volume as before. The resulting homogenatesare spun at 16,000 RPM (47,800×g Maximum, 36592×g Average) for 15minutes at 4° C. Pellets are saved and the supernatents are discarded.Pellets are combined with BUFFER to get a final volume of 300 mL, andblended using a Tekmar Tissuemizer. Initial protein content isdetermined by the Biorad protein assay. The volume of suspension isadjusted with BUFFER, such that this volume adjustment yields approx.4.0 mg/mL final concentration confirmed via Biorad protein assay. Thefinal suspension is transferred as 4.0 mL aliquots into plastic tubes,and frozen at -70 ° C.

Assay:

BUFFER is 20 mM Hepes, 1 mM EGTA, 118 mM NaCl, 5 mM KCL, 5 mM MgCl₂,0.05% BSA at pH=7.4.

Skatron filters are soaked in BUFFER with 0.1% Bovine Serum Albumin(BSA) for an hour prior to harvesting.

100 mM Bestatin and 3 mM Phosphoramidon are prepared fresh. (Final assayconcentrations will=10 mM respectively.)

Test compounds are diluted in assay binding buffer in stockconcentrations 10-fold more concentrated than desired final assayconcentrations. [¹²⁵ ]-sulfated CCK-8 labelled with Bolton-Hunterreagent (Amersham, 200 Ci/mmol) is diluted.

25 mL 100 mM Bestatin+25 mL 3 mM Phosphoramidon+25 mL test compound+50mL radioligand+25 mL buffer+100 mL guinea pig codex membranes areincubated 150 minutes at room temperature.

For B_(o) determination, assay binding buffer is substituted for testcompound.

For filter binding determination, assay buffer is substituted for testcompound and guinea pig codex membranes as well.

For non-specific binding determination, 1 mM sulphated CCK-8 (Sigma) issubstituted for test compound.

Reaction is stopped by filtering using the automated Skatron CellHarvester. The filters are rinsed using 4° C. BUFFER. The filters arepunched, placed in tubes and counted using a gamma counter.

GUINEA PIG GALL BLADDER ASSAY

Tissue Preparation:

Gall bladders are removed from guinea pigs sacrificed by cervicaldislocation. The isolated gall bladders are cleaned of adherentconnective tissue and cut into two rings from each animal (2-4 mm inlength). Rings are suspended in organ chambers containing aphysiological salt solution of the following composition (mM): NaCl(118.4); KCl (4.7); MgSO₄ ×H₂ O(1.2);CaCl₂ ×2H₂ O(2.5);KH₂ PO₄ (1.2);NaHCO₃ (25) and dextrose (11.1). The bathing solution is maintained at37° C. and aerated with 95% O₂ /5%CO₂. Tissues are connected via goldchains and stainless steel mounting wires lo to isometric forcedisplacement transducers (Grass, Model FT03 D). Responses are recordedon a polygraph (Grass, Model 7E). One tissue from each animal serves asa time/solvent control and does not receive test compound.

Assay:

Rings are gradually stretched (over a 120 min. period) to a basalresting tension of 1 gm, which is maintained throughout the experiment.During the basal tension adjustment period, the rings are exposed toacetylcholine (ACH, 10⁻⁶ M) four times to verify tissue contractility.The tissues are then exposed to a submaximal dose of sulfated CCK-8(Sigma, 3×10⁻⁹ M). After obtaining a stable response, the tissues arewashed out 3 times rapidly and every 5 to 10 minutes for 1 hour toreestablish a stable baseline.

Compounds are dissolved in dimethylsulfoxide (DMSO), then diluted withwater and assayed via a cumulative concentration-response curve to testcompound (10¹¹ to 3×10⁻⁶ M) followed by a concentration-response curveto test compound (10⁻¹¹ to 3×10⁻⁶ M) in the presence of the highest doseof the test compound. As a final test, ACH (10⁻⁶ M) is added to inducemaximal contraction. A minimum of three determinations of activity aremade for each test compound. Data for the products of Examples (1)-(19)are given in Table 1.

                  TABLE 1                                                         ______________________________________                                        In vitro Guinea Pig Gall Bladder                                              Example         % contraction (fold shift).sup.a                              ______________________________________                                        1               54.6%                                                         2               64.7%                                                         3               61.4%                                                         4               (83-fold shift)                                               5               37.9%                                                         6               56.9%                                                         7               15.6%                                                         8               51%                                                           9               (14-fold shift)                                               10              23.6%                                                         11              48.8%                                                         12              23.8%                                                         13              (125-fold shift)                                              14              (10-fold shift)                                               15              (32-fold shift)                                               16              (62-fold shift)                                               17              6%                                                            18              17.3%                                                         Intermediate i  12.2%                                                         19              (6.8-fold shift)                                              ______________________________________                                         .sup.a % acetylcholineinduced maximum contraction for the test compound a     30 mM or xfold shift of the CCK8 curve in the presence of the test            compound (30 mM).                                                        

18-HOUR DEPRIVATION-INDUCED FEEDING PARADIGM

Male, Long-Evans rats (Chades River Co., Raleigh, N.C.), weighing300-375 grams, are acclimated individually for at least a week inhanging, stainless steel mesh cages (17.8 ×25.4×17.8 cm high) with adlibitum access to water (delivered through automatic drinking spouts atthe rear of the cage) and food (Lab Blox, Purina Rodent Laboatory Chow#5001) on a 12-hour light/dark cycle (lights on from 0600-1800 hours, orh) at approximately 22.8 C. Prior to testing, all chow, but not water,is removed at 1600 h. At 0900 h the next morning, rats are weighed. At0945 h, rats are injected intraperitoneally (i.p.), orally (per os, orp.o.) or through an indwelling, intra-duodenal cannulea with a testcompound or vehicle (2 mL/kg) and returned to their home cages. Food ispresented at 1000 h. At 1030 h, remaining food and spillage is weighed.

GENERAL PROCEDURES

Unless otherwise noted, all starting materials were obtained fromcommercial suppliers and used without further purfication. The followingsolvents and reagents have been described by acronyms: tetrahydrofuran(THF), dimethylsulfoxide (DMSO), dichloromethane (DCM), trifluoroaceticacid (TFA), dimethylformamide (DMF), 1,1-carbonyldiimidazole (CDI),isobutylchloroformate (iBuCF), N-hydroxysuccinimide (HOBT),ethylcarbodiimide hydrochloride (EDC), bis(2-oxo-3-oxazolidinyl)phosphinic chloride (BOP), tert-butyloxycarbonyl (BOC), benzyloxycrbonyl(Cbz).

The ¹ HNMR spectra were recorded on either a Varian VXR-300 or a VarianUnity-300 instrument. Chemical shifts are expressed in parts per million(ppm, d units). Coupling constants are in units of hertz (Hz). Splittingpatterns are designated as s, singlet; d, doublet; t, triplet; q,quartet; m, multiplet; b, broad.

Low-resolution mass spectra (MS) were recorded on a JEOL JMS-AX505HA,JEOL SX-102 or a SCIEX-APliii spectrometers. All mass spectra were takenin the positive ion mode under electrospray ionization (ESI), chemicalionization (CI), electron impact (EI) or by fast atom bombardment (FAB)methods. Infrared (IR) spectra were obtained on a Nicolet 510 FT-IRspectrometer using a 1-mm NaCl cell. Rotations were recorded on aPerkin-Elmer 241 polarimeter. All reactions were monitored by thin-layerchromatography on 0.25-mm E. Merck silica gel plates (60F-254),visualized with UV light, 7% ethanolic phosphomolybdic acid, orp-anisldehyde solution. Flash column chromatography was performed onsilica gel (230-400 mesh, Merck).

Products were purified by preparative reversed phase-high pressureliquid lo chromatography (RP-HPLC) using a Waters Model 3000 Delta Prepequipped with a Delta-pak radial compression cartridge (C₁₈, 300 A, 15m, 47 mm×300 mm). Linear gradients were used in all cases and the flowrate was 100 mL/minute (t_(o) =5.0 min). The solvent systems used are A:water, B: 40:60, water:acetonitrile and C: acetonitrile. All solventscontained 0.1% trifluoroacetic acid (TFA). Analytical purity wasassessed by RP-HPLC using a Waters 660E system equipped with a Waters990 diode array spectrometer (t range 200-400 nM). The stationary phasewas a Vydac C₁₈ column (5 m, 4.6 mm×250 mm). The flow rate was 1.5mL/min (t_(o) =3.0 min) and the solvent systems were as described above.Data reported as tr, retention time in minutes (%B₁ to %B₂ in A overtime).

CHEMISTRY Intermediate Example A 2 -Adamantyloxychloroformate

To a 0° C. solution of 2-adamantol (5.00 g, 32.84 mmol) in CH₂ Cl₂ (82mL) was added bis(trichloromethyl) carbonate (3.61 g, 12.15 mmol),followed by pyridine (2.66 mL, 32.84 mmol) in CH₂ Cl₂ (61 mL). Thereaction mixture warmed to room temperature and stirred for two hours.The solvent was removed in vacuo at 30° C, and the residue dissolved inethyl acetate (160 mL) and stirred for 10 minutes. The pyridinium lohydrochloride was removed by filtration and the solvent removed in vacuoat 30° C. to give an oil which solidified upon standing (6.42 g, 59%);IR (CDCl₃) 1770 cm⁻¹ ; R_(f) (1:9, ethyl acetate:hexane)=0.68; ¹ HNMR(300MHz, CDCl₃) d 5.00(s, 1H), 2.18(s, 2H), 2.10-2.00(m, 2H),1.95-1.85(m, 2H), 1.8(m, 4H), 1.55-1.65(m, 2H); IR (CDCl₃) 1770 cm⁻¹.

Intermediate Example B 2-Adamantyloxycarbonyl-N-hydroxy-succinimide

To Intermediate A (3.78 g, 17.7 mmol) dissolved in chloroform (34 mL)was added N-hydroxysuccinimide dicyclohexylamine salt (5.24 g, 17.7mmol) in four equal portions. After stirring 2d, the reaction mixturewas filtered and the precipitate washed with chloroform (60 mL). Thefiltrate was washed with 10% aqueous citric acid (20 mL), 10% aqueoussodium bicarbonate (20 mL), water (20 mL), dried (MgSO₄) andconcentrated in vacuo to give the titled compound (4.09 g, 79%). R_(f)(19:1 methylene chloride:methanol)=0.74; ¹ HNMR (300 MHz, CDCl₃) d 4.89(s, 1H), 2.80 (s, 4H), 2.15 (s, 2H), 2.05-2.01 (d, J=12.9 Hz, 2H),1.89-1.84 (m, 4H), 1.80-1.72 (m, 4H), 1.64-1.55 (m, 2H).

Intermediate Example C 2-Amino-3-(1H-indol-3-yl)-2R-methyl-propionicacid methyl ester

The titled compound was prepared by the method of Roeske andAnantharamaiah, Tetrahedron Letters (1982) 23, 33, 3335-3336. Physicaland analytical data were consistent for the compound.

Intermediate Example D 2-Amino-3-(1H-indol-3-yl)-2S-methyl-propionicacid methyl ester

The titled compound was prepared by the method of Roeske andAnantharamaiah, Tetrahedron Letters (1982) 23, 33, 3335-3336. Physicaland analytical data were consistent for the titled compound.

Intermediate Example E2-[(Adamantan-2-yl-oxy)-carbonyl]-amino-3-(1H-indol-3-yl)-2RS-methyl-propionicacid

To a-methyl-(D,L) tryptophan (0.50 g, 2.29 mmol) dissolved in dioxane (5mL) was added 2-adamantyloxychloroformate (0.83 g, 2.51 mmol) and 1Naqueous NaOH (2.3 mL). The reaction mixture stirred 48h, was acidifiedwith 1N aqueous HCl and extracted with ethyl acetate (4×50 mL). Thecombined organic extracts were washed with brine (1×30 mL), dried(MgSO₄), and concentrated in vacuo to give the titled compound (1.049g). R_(f) (9:1 methanol/methylene chloride)=0.58; ¹ H NMR (300 MHz,d6-DMSO) d 8.09 (s, 1H), 7.57 (d, J=8.06 Hz, 1H), 7.32 (d, J=8.06 Hz,1H), 7.15 (t, J=7.82, Hz, 2H), 7.06(t, J=7.82, 1H), 6.98 (s, 1H), 5.29(s, 1H), 5.84 (s, 1H), 3.44 (s, 2H), 1.47- 2.06 (m, 18H); MS (FAB) m/z397 (MH⁺).

Intermediate Example F2-[(Adamantan-2-yl-oxy)-carbonyl]-amino-3-(1H-indol-3-yl)-2R-methyl-propionicacid

Step 1

To a solution of Intermediate A (0.308 g, 0.93 mmol) in dry THF (2.0 mL)was added a solution of Intermediate C (0.20 g, 0.75 mmol) in dry THF(3.5 mL) followed by a solution of triethylamine (0.21 mL, 1.50 mmol) indry THF (3.5 mL) dropwise. After 2d, the reaction mixture was dilutedwith ethyl acetate (100 mL), washed with 1N aqueous NaOH (2×30 mL), 1Naqueous HCl (2×30 mL), brine (1×30 mL), dried (MgSO₄) and concentratedin vacuo to give the methyl ester of the titled compound (0.34 g). R_(f)(9:1, methylene chloride:methanol)=0.74; ¹ HNMR (300 MHz, CDCl₃) d8.20(s, 1H), 7.58(m, 1H), 7.38(m, 1H), 7.20-7.10(m, 2H), 6.97(s, 1H),5.45(s, 1H), 4.84(m, 1H), 3.75(s, 3H), 2.20-1.40(m, 18H).

Step 2

To a solution of the ester from step 1 (0.31 g, 0.76 mmol) in aqueous1,4-dioxane (1:1) (4 mL) was added lithium hydroxide (0.048 g, 1.10mmol). The reaction mixture stirred at room temperature overnight. Thismixture was acidified with 1N aqueous HCl and was extracted into ethylacetate (6×30 mL). The combined organic extracts were washed with brine(1×30 mL), dried (MgSO₄) and concentrated in vacuo to give the titledcompound (0.22 g, 73%). R_(f) (9:1, methylene chloride:methanol)=0.33; ¹HNMR (300 MHz, CDCl₃) d 8.19(s, 1H), 7.65(d, J=8.06 Hz, 1H), 7.74-7.0(m,4H), 5.39(s,1H), 4.90(s, 1H), 3.50(s, 2H), 2.20-1.50(m, 18H).

Intermediate Example G2-[(Adamantan-2-yl-oxy)-carbonyl]-amino-3-(1H-indol-3-yl)-2S-methyl-propionicacid

Step 1

To a solution of Intermediate D (0.80 g, 3.45 mmol) in chloroform (10mL) was added Intermediate B (1.01 g, 3.45 mmol). The reaction mixturestirred overnight at room temperature. The precipitate was filtered andthe filtrate was concentrated in vacuo. The resultant residue wasdissolved in ethyl acetate (200 mL), washed with saturated aqueoussodium bicarbonate (2×30 mL), 1N aqueous HCl (2×30 mL), brine (1×30 mL),dried (MgSO₄) and concentrated in vacuo to give the methyl ester of thetitled compound (1.34 g, 76%). R_(f) (9:1, methylene chloride:methanol)=0.79.

Step 2

To a solution of the ester from Step 1 (1.34 g, 3.28 mmol) in aqueous1,4-dioxane (1:1) (50 mL) was added excess lithium hydroxide (0.61 g,14.45 mmol). The reaction mixture stirred at room temperature overnight.This mixture was partitioned between ethyl acetate (50 mL) and water (50mL) and extracted into ethyl acetate (5×50 mL). The combined organicextracts were washed with 1N aqueous HCl (2×30 mL), brine (1×30 mL),dried (MgSO₄) and concentrated in vacuo to give the titled compound(1.097 g, 80%). R_(f) (9:1, methylene chloride:methanol) =0.31; ¹ HNMR(300 MHz, d6-DMSO) d 10.91(s, 1H), 7.42(d,J=8.0 Hz, 1H), 7.30(d, J =8.0Hz, 1H), 7.02(m, 2H), 6.99(m, 1H), 4.67(s, 1H), 4.51(d, J =3 Hz, 1H),3.63(s, 1H), 3.30(obscured ABq, 2H), 2.02-1.20(m, 18H).

Intermediate Example H2R-[(Adamantan-2-yl-oxy)-carbonyl]-amino-3-(1H-indol-3-yl)-propionicacid

To a solution of Intermediate B (1.19 g, 4.06 mmol) in THF (60 mL) wasadded a o solution of D-tryptophan (0.83 g, 4.06 mmol) in 10% aqueoussodium carbonate (20 mL) and THF (10 mL). The reaction mixture stirredovernight at room temperature, was acidified with 1N aqueous HCl andextracted into ethyl acetate (6×50 mL). The combined organic extractswere washed with water (1×30 mL), brine (1×30 mL), dried (MgSO₄) andconcentrated in vacuo to give the titled compound (1.66 g, quant.).R_(f) (9:1, methylene chloride:methanol)=0.24; ¹ HNMR (300 MHz, CDCl₃) d7.56(d, J=8.0 Hz, 1H), 7.32(d, J=8.0 Hz, 1H), 7.10-6.98(m, 3H), 4.68(s,1H), 4.09(m, 1H), 3.20-3.0(m, 1H), 2.20-1.40(m, 15H).

Intermediate Example J2S-[(Adamantan-2-yl-oxy)-carbonyl]-amino-3-(1H-indol-3-yl)-propionicacid

The procedure as described for Intermediate Example H was followed,hence, L-tryptophan (1.19 g, 4.06 mmol) gave the titled product (1.53 g,94%). R_(f) (9:1, methylene chloride:methanol)=0.24; ¹ HNMR (300 MHz,CDCl₃) d 7.53(d, J=8.0 Hz, 1H), 7.30(d, J=8.0 Hz, 1H), 6.96-7.08(m, 3H),4.65(s, 1H), 4.46-4.06(m, 1H), 3.20-3.0(m, 1H), 2.20-1.40(m, 15H).

Intermediate Example K2-[(Benzyloxy)-carbonyl]-amino-3-(1H-indol-3-yl)-2RS-methyl propionicacid

To a solution of D,L-a-methyl tryptophan (5.00 g, 0.021 mol) in 10%aqueous acetone (100 mL) and sodium carbonate (2.25 g, 0.021 mol) wasadded benzyloxycarbonyl-N-hydroxysuccinimide (5.29 g, 0.021 mol) inacetone (50 mL). The solution was stirred overnight. The pH was adjustedto 4 with 1N aqueous HCl, and the solution was concentrated in vacuo.The residue was partitioned between ethyl acetate (500 mL) and aqueous1N HCl (300 mL), and the organic layer was separated, washed with water(500 mL), dried (Na₂ SO₄) and concentrated in vacuo to give a whitesolid (6.9 g, 0.019 mol) which was used without further purification.R_(f) (9:1, methylene chloride:methanol)=0.16; MS (FAB) m/e 353 (MH⁺).

Intermediate Example L2-[(t-Butyloxy)-carbonyl]-amino-3-(1H-indol-3-yl)-2RS-methyl-propionicacid

Intermediate L was prepared analogously to Intermediate E. Hence toa-methy-D,L-tryptophan (2.18 g, 10.0 mmol) was addeddi-tert-butyldicarbonate (2.40 g, 11.0 mmol) to give the titled product(2.25 g, 76%). R_(f) (9:1, methylene chloride:methanol)=0.24; ¹ HNMR(300 MHz, CDCl₃) d 7.45(d, J=8.0 Hz, 1H), 7.30(d, J =8.0 Hz, 1H),7.02-6.90(m, 3H), 3.10(obscured ABq, 2H), 1.38(s, 9H), 1.25(s, 3H).

Intermediate Example M 2-[(t-Butyloxy)-carbonyl]-amino-3-(1H-indol-3-yl)-2R-methyl-propionic acid

Intermediate M was prepared analogously to Intermediate E. Hence,Intermediate C (227.0 mg, 0.85 mmol) gave the titled product. R_(f)(9:1, methylene chloride:methanol) =0.24.

Intermediate Example NN-Adamantan-2-yl-2-amino-3-(1H-indol-3-yl)-2RS-methyl-propionicamide

Step 1

To a solution of Intermediate K (0.352 g, 0.001 mol), 2-aminoadamatane(0.187 g, 0.001 mol), N-hydroxybenztriazole (0.135 g, 0.001 mol), andtriethylamine (0.140 mL, 0.001 moles) in dichloromethane (35 mL) wasadded EDC (0.191 g, 0.001 mol). The solution was allowed to stir at roomtemperature overnight. The reaction mixture was diluted withdichloromethane (200 mL) and extracted successively with 1N aqueous HCl(100 mL), water (100 mL), 10% aqueous sodium bicarbonate (100 mL) andwater (100 mL), then dried (Na₂ SO₄) and evaporated to give the crudeamide (0.390 g, 0.75 mmol) which was used without further purification.R_(f) (9:1, methylene chloride:methanol)=0.78; MS (FAB) 486 (MH⁺).

Step 2

The amide from step 2 (0.39 g, 0.75 mmol) was dissolved in MeOH (5 mL)and glacial acetic acid (1 mL) and 10% Pd/C (50 mg) was added. Themixture was shaken at 50 psi until the reaction was judged complete byTLC. The mixture was filtered through a celite pad, and the pad waswashed with methanol (100 mL). The methanol was evaporated in vacuo andthe residue partitioned between ethyl acetate (50 mL) and 1N aqueousNaOH (50 mL). The organic layer was separated, washed with water (100mL), dried (Na₂ SO₄) and evaporated to give the product as a white foam(0.27 g, 0.7 mmol) which was used without further purification. R_(f)(9:1, methylene chloride:methanol)=0.18; MS (FAB) m/e 352 (MH⁺).

Intermediate Example O1-(6-Amino-5R-[(t-Butyloxy)carbonyl]-amino-hexyl)-3-o-tolyl-urea

Step 1

To N-a-BOC-D-lysine (5.0 g, 20.48 mmol) dissolved in 1N aqueous sodiumhydroxide (20.48 mmol) was added o-toluylisocyanate (2.53 mL, 20.48mmol). The reaction mixture stirred 3h, was acidified with 1N aqueousHCl and extracted into ethyl acetate 5 (6×50 mL). The combined organicextracts were washed with brine (1×30 mL), dried (MgSO₄) andconcentrated in vacuo to give 7.93 g of crude urea. R_(f) (9:1,methanol:methylene chloride)=0.06; ¹ H NMR (300 MHz, d6-DMSO) d 7.81 (d,J=8.06 Hz, 1H), 7.58 (s, 1H) 7.08 (m, 3H), 6.83 (t, J=7.33 Hz, 1H), 6.51(t, J=4.88 Hz, 1H), 3.83 (m, 1H), 3.05 (d, J=5.13 Hz, 2H), 2.15 (s, 3H),1.36 (s, 13H); MS (El) m/z 380 (MH⁺).

Step 2

To the crude urea from step 1 (1.98 g, 5.22 mmol) dissolved in THF (47mL) was added 1,1-carbonyl-diimidazole (1.69 g, 10.4 mmol). The reactionmixture stirred 16h at room temperature. This mixture was cooled to 0°C. and a solution of lithium aluminum hydride (26.1 mL of a 1M THFsolution) was added dropwise. After 2h 2N aqueous NaOH was addeddropwise and the aqueous layer was extracted with ethyl acetate (4×50mL). The combined organic extracts were washed with 1N aqueous HCl (1×30mL), brine (1×30 mL), dried (MgSO₄) and concentrated in vacuo to give(1.00 g, 52%) of the product alcohol: R_(f) (9:1, methanol:methylenechloride)=0.60; ¹ HNMR (300 MHz, d6-DMSO) d 7.79 (d, J=8.1 Hz, 1H), 7.52(s, 1H), 7.01-7.08 (m,2H), 6.79-6.84 (m,1H), 6.42-6.50 (m, 2H), 4.54 (m,1H), 3.15-3.29 (m, 2H), (m, 2H), 2.13 (s, 3H), 1.34 (s, 9H), 1.29-1.52(m, 6H); MS (FAB) miz 366 (MH⁺).

Alternative procedure for formation of 2

To the crude urea from step 1 (7.9 g, 20.9 mmol) in THF (210 mL) cooledto 0° C. was added isobutyl chloroformate (3.3 mL, 25.1 mmol) anddiisopropylethylamine (4.5 mL, 25.1 mmol). The reaction mixture stirred1h at 0° C. and then was added to a THF (210 mL) slurry of lithiumaluminum hydride (1.59 g). The reaction stirred 48h at room lotemperature, was acidified with 1N aqueous HCl and diluted with ethylacetate (500 mL). The reaction mixture was filtered through a pad ofcelite and the filtrate was extracted with ethyl acetate (3×100 mL)washed with brine (1×50 mL), dried (MgSO₄) and concentrated in vacuo.Purification by flash chromatography (SiO₂ 1.1, hexanes:ethyl acetate)gave (4.18 g, 55%) of the product alcohol.

Step 3

To a solution of the product alcohol of step 2 (4.00 g, 10.96 mmol) andtriethylamine (1.83 mL, 13.15 mmol) dissolved in methylene chloride (22mL) and cooled to 0° C. was added methanesulphonyl chloride (1.02 mL,13.15 mmol) dropwise. The reaction mixture stirred at room temperatureovernight. The resultant mixture was diluted with methylene chloride,washed with aqueous citric acid (2×30 mL), water (1×30 mL), brine (1×30mL), dried (MgSO₄), and concentrated in vacuo. Purification by stepwiseflash chromatography (SiO₂ 50% hexanes:ethyl acetate, then 30%hexanes:ethyl acetate) gave 1.78 g of the product mesylate: R_(f) (1:1hexanes:ethyl acetate),=0.16 (9:1, methanol:methylene chloride)=0.55; ¹HNMR (300 MHz, d6-DMSO) d 7.79 (d, J=8.1 Hz, 1H), 7.54 (s, 1H),7.01-7.09 (m, 2H), 6.80-6.91 (m, 2H), 6 50- 651 (m,1H), 3.99-4.10 (m,2H), 3.63 (s, 1H), 3.13 (s, 3H), 3.04-3.07 (m, 2H), 1.33 (s, 9H),1.25-1.49 (m, 6H); MS (FAB) m/z 444 (MH⁺).

Step 4

To the product mesylate of step 3 (1.78 g, 4.0 mmol) dissolved indimethylformamide (40 mL) was added sodium azide (0.52 g, 8.00 mmol).The reaction mixture was heated at 80° C. for 2h, then poured into icewater and extracted with ethyl acetate (3×50 mL). The combined organicextracts were washed with water (3×30 mL), dried (MgSO₄) andconcentrated in vacuo. Purification by flash chromatography (SiO₂, 1:1,hexanes:ethyl acetate) gave (0.68 g, 44%) of the product azide: R_(f)(1:1, hexanes:ethyl acetate)=0.2; ¹ HNMR (300 MHz, d6-DMSO) d 7.79 (d,J=7.8 Hz, 1H), 7.53 (s, 1H), 7.02-7.09 (m, 2H), 6.80-6.88 (m, 2H), 6.49(m, 1H), 3.52 (s, 1H), 3.22 (d, J=5.9 Hz, 2H), 3.04 (m, 2H), 2.14 (s,3H), 1.23-1.43 (m, 6H), 1.34 (s, 9H); MS (FAB) m/z 391 (MH⁺).

Step 5

To the product azide of step 4 (0.68 g, 1.7 mmol) dissolved in ethanol(10 mL) was added 10% palladium on carbon (68 mg). The reaction mixturewas stirred under 1 atmosphere of hydrogen for 12h, filtered through apad of celite and concentrated in vacuo. Purification by flashchromatography (SiO₂, 1:9, methanol:methylene chloride) gave (0.59 g,95%) of product amine: R_(f) (1:9, methanol:methylene chloride)=0.15; ¹HNMR (300 MHz, d6-DMSO) d 7.79 (d, J=8.06 Hz, 1H), 7.53 (s, 1H),7.02-7.09 (m, 2H), 6.82 (t, J=7.3 Hz, 1H), 6.48-6.50 (m, 2H), 3.31 (s,3H), 3.0 (m, 2H), 243 (d, J=6.1 Hz, 2H), 2.14 (s, 3H), 1.35 (s, 9H),1.23-1.56 (m, 6 H); MS (ESI) m/z 365 (MH⁺).

Intermediate Example P

1-(6-Amino-5S-[(t-Butyloxy)carbonyl]-amino-hexyl)-3-o-tolyl-urea

Intermediate P was prepared analogously to Intermediate O. Hence,N-a-t-butyloxycarbonyl-L-lysine (5.42 g, 14.3 mmol) gave theenantiomeric compound (115.0 mg, 2%).

Intermediate Example QN-6-[(Benzyloxy)carbonyl]-N-2-[(t-Butyloxy)carbonyl-hexane-1,2R,6-triamine

Intermediate Q was prepared analogously to Intermediate 0. Hence,N-a-t-butyloxycarbonyl-N-e-benzyloxycarbonyl-D-lysine (4.90 g, 12.9mmol) gave the product (1.31 g, 28%). R_(f) (9:1, methylenechloride:methanol)=0.2; ¹ HNMR (300 MHz, d6 DMSO) d 7.25-7.37(m, 5H),7.17-7.20(m, 1H), 6.46(d, J=8.5 Hz, 1H), 4.97(s, 2H), 2.97-2.91(m, 2H),2.42(d, J =5.9 Hz, 2H), 1.35(s, 9H), 1.41-1.16(m,6H). (In CDCl₃ broadpeak seen at 3.53 ppm for methine); MS (ESI) m/z 366 (MH⁺)

Intermediate Example R3R-[(t-Butyloxy)carbonyl]-amino-7-(3-o-tolyl-ureido)-heptanoic acid

Step 1

To a solution of2-[(t-butyloxy)carbonyl]-amino-6-(3-o-tolyl-ureido)-hexanoic acid (3.8g, 0.01 mol) and N-methylmorpholine (1.1 mL, 0.01 mol) intetrahydrofuran (100 mL) at -15° C. was added isobutylchloroformate (1.3mL, 0.01 mol) in tetrahydrofuran (20 mL). The mixture was stirred 1h at0° C., whereupon a solution of freshly prepared diazomethane in etherwas added until a persistent yellow color was obtained. The resultantsolution was allowed to stir overnight, then the residual diazomethanewas quenched by the addition of glacial acetic acid. The tetrahydrofuranwas evaporated in vacuo and the residual yellow oil was partitionedbetween ethyl acetate (300 mL) and saturated aqueous NaHCO₃ (300 mL).The organic layer was separated, washed with water (300 mL), dried(MgSO₄) and concentrated in vacuo. The crude diazomethylketone (3.5 g)was purified by chromatography over silica gel (hexane:ethyl acetate,1:1) to give 1.0 g of a bright yellow solid. R_(f) (3:1, ethylacetate:hexane)=0.33; ¹ HNMR (CD₃ OD):delta=6.1 (1H, CH═N₂); MS (FAB)m/e 404 (MH⁺).

Step 2

The diazomethylketone from step 1 (1.9 g, 4.7 mmol) was dissolved inMeOH (50 mL) and silver benzoate (0.072 g, 4.7 mmol) in triethylamine (1mL) was added. After the evolution of N₂ was complete, a second portionof silver benzoate was added, and the solution was allowed to stir for 1h at room temperature, whereupon activated charcoal (0.5 gm) was added,and the solution was filtered by gravity. The filtrate was concentratedin vacuo, and partitioned between ethyl acetate (300 mL) and 10% aqueouscitric acid (300 mL). The organic layer was separated, washed with water(300 mL), dried (Na₂ SO₄) and concentrated in vacuo to give 1.84 gramsof the beta-amino ester which was used in the following step withoutfurther purification. R_(f) (3:1, ethyl acetate:hexane)=0.44; MS (FAB)m/e 408 (MH⁺); NMR (d6-DMSO): delta=3.62 (s, 3H, CH₂ COOCH₃) 2.43 (d,2H, CH₂ COOCH₃)

Step 3

The b-amino ester from step 2 (1.84 g, 4.7 mmol) was dissolved inmethanol (50 mL) and water (2 mL) and 6N aqueous NaOH (0.8 mL) wasadded. The solution was allowed to stir overnight, then neutralized with1N HCl, and concentrated in vacuo. The residual oil was partitionedbetween 10% aqueous sodium bicarbonate (300 mL) and ethyl acetate (300mL). The organic layer was separated and washed with water (300 mL). Theaqueous layer was extracted twice with ethyl acetate (300 mL). Theorganic layers were combined, washed with water (300 mL), dried (Na₂SO₄) and concentrated in vacuo to give 1.7 g of the product b-amino acidas an amorphous solid. R_(f) (250:25:1, methylene chloride:methanol:acetic acid)=0.29; MS (FAB): m/e =394 (MH⁺).

Intermediate Example S 1-(6-Amino-5S-[(benzyloxy)carbonyl]-aminohexyl)-3-o-tolyl-urea

Step 1

To aqueous sodium hyroxide (36.0 mL, 36.0 mmol) was added toN-a-Cbz-lysine (10.00 g, 36.0 mmol). The resultant mixture stirred atroom temperature for 5 min prior to the addition of o-tolylisocyanate(3.90 mL, 36.0 mmol). The resultant mixture was stirred at roomtemperature for 18h. During this time the reaction mixture turned solid.The resultant solid was dissolved in water (50 mL) and the pH wasadjusted to 2.0 with concentrated hydrochloric acid to precipitate aflocculant white gum. The product was extracted into ethyl acetate (3×60mL). The resultant organic extracts were washed with water (2×50 mL),dried (MgSO₄) and concentrated in vacuo to afford product as a whiteglassy solid (14.0 g). [a]_(D) (c=0.0152, methanol)=-2.69° ; ¹ HNMR (300MHz, d6-DMSO) d 7.8(d,1H), 7.60(d, 2H), 7.38(b, 5H), 7.08(m, 2H),6.82(t, 1H), 650(t, 1H), 5.0(s, 2H), 3.03(b, 2H), 2.18(s, 3H),1.5-1.8(m, 2H), 1.40(b, 4H).

Step 2

A solution of the above compound (9.50 g, 22.8 mmol) and1,1-carbonyl-diimidazole (5.16 g, 34.2 mmol) was stirred at roomtemperature for 16 h. Sodium borohydride (4.14 g, 114.0 mmol) was addedand the resultant mixture stirred vigorously at room temperature for 1.5h prior to the addition of a further batch (1.00 g) of sodiumborohydride. The resultant mixture was stirred vigorously at roomtemperature for a further 3h, then neutralized with 2N aqueoushydrochloric acid pH6. The resultant mixture was extracted into ethylacetate (3×100 mL), the combined organic extracts were washed with 2Naqueous sodium hydroxide (2×50 mL), water (50 mL), brine (50 mL), dried(MgSO₄) and concentrated in vacuo to afford the product as a white solid(7.40 g). [a]_(D) (c=0.0026, methanol)=-8.88°; ¹ HNMR (300MHz, d6-DMSO)d 7.8(d, 1H), 7.58(s, 1H), 7.3a(b, 5H), 7.68(m, 2H), 6.83(t, 1H),6.51(t, 1H), 4.98(s,2H), 4.60(b, 1H), 3.2-3.4(b, 2H), 2.1a(s, 3H),1.2-1.6(b, 6H);

Step 3

To a solution of the product above (7.10 g, 17.77 mmol), indichloromethane (100 mL) was added triethylamine (3.69 mL, 26.52 mmol),tosylchloride (3.39 g, 17.75 mmol) and N,N-dimethylaminopyridine (20mg). The resultant reaction mixture was stirred at room temperature for16h, washed with 2N aqueous sodium hydrogen carbonate (100 mL), 0.5Naqueous citric acid (50 mL), and water (50 mL) prior to drying (MgSO₄)and concentration in vacuo to afford the product (8.84 g) as a whitefoam which was used crude in the next reaction. R_(f) (9:1 methylenechloride:methanol)=0.52; ¹ HNMR (300MHz, CDCl₃) d 7.80(d, 2H),7.10-7.40(m, 15H), 6.03(s, 1H), 5.03(s, 2H), 4.01(d, 2H), 3.80(b, 1H),3.20(b, 2H), 2.41 (s, 3H), 2.21 (s, 3H), 1-2-1.6 (m, 6H).

Step 4

A mixture of the product from step 3 (8.84 g, 15.96 mmol) and sodiumazide (2.06 g, 31.98 mmol) in N,N-dimethylformamide (120 mL) was heatedat 80° C. for 1.25 h. The reaction mixture was cooled, poured into water(150 mL) and extracted into ethyl acetate (3×100 mL). The combinedorganics were washed with water (100 mL), brine (100 mL), dried (MgSO₄)and concentrated in vacuo to afford the product (6.70 g) as a white foamwhich was used crude in the next reaction. R_(f) (2:1,ethylacetate:hexane)=0.48; ¹ HNMR (300 MHz, CDCl₃) d 7.05-7.4(b, 11H),6.02(s, 1H), 5.03(s, 2H), 3.40(b, 2H), 3.20(b, 2H), 2.21 (s, 3H),1.2-1.6(m, 6H).

Step 5

A mixture of the product above (6.70 g, 15.58 mmol), Lindlar catalyst(2.68 g) and ethyl acetate (150 mL) was shaken under 50 psi hydrogenusing a Parr hydrogenation apparatus. After 5h the solids were removedby filtration through a pad of celite, the filtrate concentrated invacuo and the resultant residue partitioned between dichloromethane (100mL) and 2N aqueous hydrochloric acid (3×50 mL). The combined aqueousphase was then adjusted to pH7 with solid sodium hydrogen carbonate andthen to pH10 with 2N aqueous sodium hydroxide and extracted intomethylene choloride (3×80 mL). The combined organic extracts were dried(MgSO₄ /K₂ CO₃) and concentrated in vacuo to afford the product (900 mg)as a cream solid. A solid that precipitated from the aqueous phase wasisolated by filtration and dried by azeotrope with acetonitrile toafford additional product (710 mg). The titled product was used crude inthe next reaction. R_(f) (9:1:0.2, methylenechloride:methanol:triethylamine)=0.31; ¹ HMNR (300 MHz, CDCl₃) d 7.59(d,2H), 7.0- 7.4(m, 8H), 6.58(s, 1H), 5.31 (b, 2H), 5.03(s, 2H), 3.6(b,1H), 3.20(b, 2H), 2.62(ddd, 2H), 2.21 (s, 3H), 1.2-1-6(m, 6H).

Intermediate Example T 1-(6-Amino-5R-[(benzyloxy)carbonyl]-aminohexyl)-3-o-tolyl-urea

This was prepared in a manner analogous to intermediate S. Hence,N-a-Cbz-D-lysine (10.00 g, 36 mmol) gave the product (1.00 g) as an offwhite solid. R_(f) (9:1:0.2, methylenechloride:methanol:triethylamine)=0.31; ¹ HNMR (300 MHz, d6 DMSO) d7.82(d, 1H), 7.63(s, 1H), 7.3-7.4(m, 4H), 7.03(m, 2H), 6.81(t, 1H),6.62(t, 1H), 5.0(apparent q, 2H), 3.60(b, 2H), 3.02(b, 2H), 2.18(s, 3H),1.2-1.6(b, 6H); [a]_(D) (c=0.01285, MeOH)=+2.68°; MS (ESI) m/z, 389(MH⁺).

Intermediate Example U 1-(5-Amino-4R-[(benzyloxy)carbonyl]-aminopentyl)-3-o-tolyl-urea

Step 1

A solution of N-d-BOC-D-ornithine (4.0 g, 10.9 mmol) in 4N HCl indioxane (40 mL) was stirred at room temperature for 1.5 h. The solidsthat formed were removed by filtration, washed with ether and driedunder vacuum to afford the product (3.11 g) as a white powder. ¹ HNMR(300 MHz, d6-DMSO) d 8.0(b, 3H), 7.42(d, 1H), 7.38(s, 5H), 3.98(b, 1H),2.78(b, 2H), 1.5-1.8(m, 2H).

Step 2

This was prepared as for step 1 of intermediate S. Hence the materialfrom step 1 (3.11 g, 10.28 mmol) gave the product (3.48 g) as a whitesolid. ¹ HNMR (300 MHz, d6-DMSO) d 7.8(d, 1H), 7.60(d, 1H), 7.56(s, 1H),7.38(b, H), 7.06(m, 2H), 6.83(t, 1H), 6.50(t, 1H), 5.01(s, 2H), 3.95(dt,1H), 3.02(b, 2H), 2.10(s, 3H), 1.4-1.8(b, 4H).

Step 3

This was prepared in a similar manner to step 2 of intermediate S. Hencethe material from step 2 (3.48 g, 8.72 mmol) gave the product (3.23 g)as a white solid. R_(f) (9:1:0.1, methylenechloride:methanol:triethylamine)=0.78; ¹ HNMR (300 MHz, CDCl₃ +d6 DMSO,20:1) d 7.61(d, 2H), 7.3(b, 5H), 7.10(m, 2H), 6.90(m, 3H), 5.8(b, 1H),5.75(d, 1H), 5.0(s, 2H), 3.16(b, 2H), 2.19(s, 3H), 1.4-1.5(b, 4H); MS(ESI) m/z 386.4 (MH⁺).

Step 4

This was prepared in a similar manner to step 3 of intermediate S. Hencethe material from step 3 (3.22 g, 8.35 mmol) gave the product (4.25 g)as a cream foam that was used crude in the next reaction. R_(f) (9:1methylene chloride:methanol)=0.51; ¹ HNMR (300 MHz, d6-DMSO) d 7.78(m,3H), 7.55(s, 1H), 7.40(d, 2H), 7.36(b, 6H), 7.04(m, 2H), 6.82(t, 1H),6.50(t, 1H), 4.98(s, 2H), 3.96(d, 2H), 3.04(b, 1H), 3.0(b, 2H), 2.40(s,3H), 2.18(s, 3H), 1.40(b, 4H).

Step 5

This was prepared in a similar manner to step 4 of intermediate S. Hencethe material from step 4 (4.25 g, 7.88 mmol) gave the product (2.51 g)as a white powder which was used crude in the next step. R_(f) (2:1ethyl acetate:hexane)=0.47; ¹ HNMR (300 MHz, d6-DMSO) d 7.8(d, 1H),7.55(s, 1H), 7.38(b, 6H), 7.05(m, 2H), 6.82(t, 1H), 6.50(t, 1H), 5.02(s,2H), 3.61 (b, 1H), 3.25(b, 2H), 3.03(b, 2H), 2.15(s, 3H), 1.40(b, 4H).

Step 6

This was prepared in a similar manner to step 5 of intermediate S exceptthat tetrahydrofuran was the sovent. Hence the material from step 5(1.25 g) gave the product (693 mg) as a white solid. R_(f) (9:1:0.1,methylene chloride:methanol:triethylamine) =0.22; ¹ HNMR (300 MHz,d6-DMSO) d 7.80(d, 1H), 7.62(s, 1H), 7.38(b, 1H), 7.06(apparent q, 3H),6.84(t, 1H), 6.60(b, 1H), 5.0(s, 2H), 3.25(b, 2H), 2.15(s, 3H), 1.40(b,4H); MS (ESI) m/z 385.4 (MH⁺).

Intermediate Example V1-(5-Amino-4S-[(benzyloxy)carbonyl]-amino-pentyl)-3-o-tolyl-urea

This was prepared in a similar manner to intermediate U. HenceN-a-Cbz-L-ornithine (5.00 g, 18.7 mmol) gave the product (1.31 g) as awhite glassy foam after flash column chromatography (95:5:0.1, methylenechloride:triethylamine). R_(f) (1% Et₃ N, 10% MeOH, CH₂ Cl₂)=0.22; ¹HNMR (300 MHz, d6-DMSO) d 7.80(d, 1H), 7.60(s, 1H), 7.38(b, 6H), 7.06(m,3H), 6.84(t, 1H), 6.60(b, 1H), 5.0(s, 2H), 3.25(b,2H), 3.0(d,2H),2.15(s, 3H), 1.40(b, 4H); MS (ESI) m/z 385.4 (MH⁺).

Intermediate Example W1-(5-Amino-4S-[(benzyloxy)carbonyl]-amino-pentyl)-3-o-tolyl-urea

Step 1

This was prepared in a similar manner to step 1 of intermediate S. HenceN-a-BOC-L-ornithine (5.0 g, 21.55 mmol) gave the product (5.63 g) as awhite foam. ¹ HNMR (300 MHz, d6-DMSO) d 7.8(d, 1H), 7.58(s, 1H), 7.08(m,2H), 6.82(t, 1H), 6.50(t, 1H), 3.83(dt, 1H), 3.02(m, 2H), 2.17(s, 3H),1.4-1.7(m, 4H), 1.30(s, 9H); MS (ESI) m/z 729.2 (MH)⁺ ₂.

Step 2

This was prepared in a similar manner to step 2 of intermediate S. Hencethe material from step 1 (5.63 g, 15.40 mmol) gave the product (3.34 g)as a white solid after flash column chromatography (95:5, methylenechloride:methanol). R_(f) (9:1, methylene chloride:methanol)=0.27; ¹HNMR (300 MHz, d6-DMSO) d 7.8(d, 1H), 7.58(s, 1H), 7.04(m, 2H), 6.81(b,2H), 4.60(b, 1H), 3.25(b, 2H), 2.15(s, 3H), 1.2-1.6(c+s, 13H).

Step 3

This was prepared in a similar manner step 3 of intermediate S. Hencethe above product (2.84 g, 8.09 mmol) gave the titled product (3.08 g,80% pure). This mixture was used crude in the following reaction. R_(f)(9:1, methylene chloride:methanol)=0.52; ¹ HNMR (300 MHz, d6-DMSO) d7.80(m, 3H), 7.50 (s, 1H), 7.44(d, 1H), 7.08(m, 3H), 6.82(m, 2H),6.48(m, 2H), 3.90(b, 2H), 3.60(b, 1H), 3.0(b, 2H), 2.50(s, 3H), 2.10(s,3H), 1.1-1.4(m, 13H).

Step 4

This was prepared in a similar manner to step 4 from intermediate S.Hence the product from step 3 (3.08 g, 6.09 mmol) gave the product (940mg) as a colorless glass which contained about 10% impurities. Thismaterial was used crude in the following reaction. R_(f) (9:1, methylenechloride:methanol)=0.50; ¹ HNMR (300 MHz, d6-DMSO) d 7.80(d, 1H), 7.50(s, 1H), 7.05 (m, 2H), 6.81(t, 1H), 6.50(b, 2H), 3(b, 1H), 3.22(2dd,2H), 3.03(b, 2H), 2.08(s, 3H), 1.1-1.6(m, 13H); MS (ESI) m/z 377.4(MH⁺).

Step 5

A mixture of the above compound (940 mg) and 10% palladium on carbon (95mg) in ethanol (50 mL) was stirred under an atmosphere of hydrogen for20 h. The solids were removed by filtration through celite and thefiltrate was concentrated in vacuo. The residue was partitioned betweenethyl acetate (50 mL) and 10% aqueous citric acid (3×50 mL). Thecombined aqueous extracts were washed with ethyl acetate (50 mL), the pHadjusted to pH11 (2N aqueous sodium hydroxide) and extracted into ethylacetate. The latter combined organic extracts were washed with water,brine, dried (K₂ CO₃) and concentrated in vacuo. The residue waspurified by flash column chromatography (9:1:0.1, methylenechloride:methanol:ammonia) to afford the product (80 mg) as a clearcolorless foam. R_(f) (9:1:9.1, methylene chloride:methanol:ammonia)=0.22; ¹ HNMR (300 MHz, CDCl₃) d 7.50(b, 1H), 7.20 (b, 3H), 7.03 (b,1H), 6.40(s, 1H), 5.30(b, 1H), 4.80(d, 1H), 3.5e(b, 1H), 3.20(b, 2H),2.62(2dd, 2H), 2.22(s,3H), 1.1-1.5(mm, 13H); MS (ESI) m/z 351.4 (MH⁺).

EXAMPLE 1N-[1R-[2-Adamantan-2-yl-oxy)carbonyl]-amino-3-(1H-indol-3-yl)-2RS-methyl-propionylamino-methyl]-5-(3-o-tolyl-ureido)-pentyl]-succinamicacid

Step 1

To Intermediate O (0.17 g, 0.47 mmol) dissolved in dimethylformamide (2mL) and methylene chloride (2 mL) was added 1-hydroxybenztriazole (0.19g, 1.40 mmol), Intermediate E (0.19 g, 0.93 mmol), and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.22 g,1.12 mmol). The reaction mixture stirred at room temperature for 24h,was diluted with ethyl acetate, extracted with saturated aqueous NaHCO₃(2×30 mL), 1N aqueous HCl (2×30 mL), brine (1×30 mL), dried (MgSO₄), andconcentrated in vacuo to give the coupled product (0.53 g).

Step 2

The product of step 1 was dissolved in 4N HCl in dioxane (5 mL) andstirred for 1 h at room temperature. The reaction mixture formed a gumon addition of ethyl ether. The ethyl ether was decanted and the gumtriturated with ethyl ether (3×30 mL) to give 0.22 g of the aminehydrochloride salt (0.32 mmol, 69% 2-steps); R_(f) (9:1, methylenechloride:methanol)=0.14; ¹ HNMR (300 MHz, CD₃ OD) d 7.40 (t, J=8.32 Hz,2H), 7.23 (d, J=7.82 Hz, 1H), 6.88-7.06 (m, 6H), 3.00-3.31 (m, 5H), 2.15(s, 3H), 1.30 1.94(m, 24H); MS (FAB) m/z 643 (MH⁺).

Step 3

To the product of step 2 (0.20 g, 0.31 mmol) dissolved indimethylformamide (0.62 mL) was added triethylamine (86 mL, 0.62 mmol)and succinic anhydride (40 mg, 0.40 mmol). The reaction mixture stirredovernight, was diluted with MeOH/CH3CN/H₂ O and purified by preparativeRP-HPLC chromatography (60-90%B in A, 30 min) to give the product (15.3mg, 7%). The diastereomers were >98% pure and had minimal separation asdetermined on analytical RP-HPLC: tr=19.0 min (70-100%B in A, 30 min).MS (FAB) m/z 743 (MH⁺).

EXAMPLE 2N-[1R-[2-Adamantan-2-yl-oxy)carbonyl-amino-3-(1H-indol-3-yl)-2R-methyl-propionylamino-methyl]-5-(3-o-tolyl-ureido)-pentyl]-succinamic acid

Example 2 was prepared analogously to Example 1. Hence Intermediate F(200.0 mg, 0.05 mmol) was coupled to Intermediate 0 (182.0 mg, 0.05mmol) to give the product (69.4 mg, 19%) after preparative RP-HPLCpurification (60-90% B, 30 min). The lyophile was >98% pure asdetermined on analytical RP-HPLC: tr=28.9 min (60-90%B in A, 30 min).R_(f) (10:1:0.1, methylene chloride:methanol:acetic acid)=0.27; ¹ HNMR(300 MHz, CD₃ OD) d 7.50(d, J=8.0 Hz, 2H), 7.31 (d, J=8.0 Hz, 1H),7.20-6.90(m, 6H), 3.88(s, 1H), 3.50(m, 2H), 3.21 (m, 3H), 3.0(m, 1H),2.58-2.45(m, 4H), 2.23(s, 3H), 2.02-1.38(m, 24H); MS (ESI) m/z 743 (MH⁺).

EXAMPLE 3N-[1R-[2-Adamantan-2-yl-oxy)carbonyl]-amino-3-(1H-indol-3-yl)-2S-methyl-propionylamino-methyl]-5-(3-o-tolyl-ureido),pentyl]-succinamicacid

Example 3 was prepared analogously to Example 1. Hence, Intermediate O(182.0 mg, 0.50 mmol) was coupled to Intermediate G (200.0 mg, 0.50mmol) to give the product (64.8 mg, 17%) after preparative RP-HPLCpurification (60-90% B in A, 30 min). The lyophile was >98% pure asdetermined on analytical RP-HPLC: tr=29.73 (60-90%B, 30 min). R_(f)(10:1:0.1, methylene chloride:methanol:acetic acid)=0.27; ¹ HNMR (300MHz, CD₃ OD) d 7.50 (d, J=8.0 Hz, 2H), 7.31 (d, J=8.0 Hz, 1H), 7.20-6.90(m, 6H), 3.88 (s, 1H), 3.38 (m, 2H), 3.19-3.0 (m, 4H), 2.58 (m, 24H); MS(ESI) m/z 743 (MH⁺).

EXAMPLE 4 N-[1R-[2S-Adamantan-2-yl-oxy)carbonyl-amino-3-(1H-indol-3-yl)-propionylamino-methyl-5-(3-o-tolyl-ureido)-pentyl]-succinamicacid

Example 4 was prepared analogously to Example 1. Hence, Intermediate O(100 mg, 0.27 mmol) was coupled to Intermediate J (105 mg, 0.27 mmol) togive the product (92.7 mg, 0.127 mmol, 46%) after RP-HPLC purification(60-80% B - 30 min). The lyophile was >98% pure as determined onanalytical RP-HPLC: tr =21.60 (60-80% B in A, 30 min). ¹ HNMR (300 MHz,CD₃ OD) d 7.52(d, J=7.6 Hz, 1H), 7.41 (d, J=7.8 Hz, 1H), 7.24(d, J=7.8Hz, 1H), 6.88-7.08(m, 6H), 4.59(b, 1H), 4.30-4.32(m, 1H), 3.76(s, 1H),2.94-3.18(m, 6H), 2.46-2.48(s, 2H), 2.24-2.32(m, 2H), 2.14(s, 3H),1.08-2.01 (m, 20H); MS (ESI) m/z 729 (MH⁺).

EXAMPLE 5N-[1R-[2R-Adamantan-2-yl-oxy)carbonyl]-amino-3-(1H-indol-3-yl)-propionylamino-methyl]-5-(3-o-tolyl-ureido)-pentyl]-succinamic acid

Example 5 was prepared analogously to Example 1. Hence, Intermediate O(100 mg, 0.274 mmol) was coupled to Intermediate H (105 mg, 0.274 mmol)to give the product (75.8 mg, 0.104 mmol, 38%) after RP-HPLCpurification (60-80% B in A, 30 min). The lyophile was >98% pure asdetermined on analytical RP-HPLC: tr=20.1 (60-80% B in A, 30 min). ¹HNMR (300 MHz, CD₃ OD) d 7.51 (d, J=7.6 Hz, 1H), 7.42(d, J=7.8 Hz, 1H),7.23(d, J=7.8 Hz, 1H), 6.91-7.08(m, 6H), 4.59-4.65(m, 1H), 4.30-4.32(m,1H), 3.72-3.73(m, 1H), 3.00-3.16(m, 6H), 2.47-2.49(m, 2H), 2.31-3.34(m,2H), 2.15(s, 3H) 1.27-1.97(m, 20 H); MS (ESt) m/z 729 (MH⁺).

EXAMPLE 6N-[1S,[2-Adamantan-2-yl-oxy)carbonyl]-amino-3-(1H-indol-3-yl)-2RS-methyl-propionylamino-methyl]-5-(3-o-tolyl-ureido)-pentyl]-succinamicacid

Step 1

A solution of Intermediate E (100 mg, 0.252 mmol), EDC (490 mg, 0.252mmol) Intermediate S (100 mg, 0.252 mmoi) and HOBT (34.0 mg, 0.252 mmol)in methylene chloride (5 mL) was stirred at room temperature for 16 h,The solvent was removed in vacuo and the residue dissolved in ethylacetate (10 mL). The organic layer was washed with 2N aqueoushydrochloric acid (10 mL), 2N sodium hydroxide solution (10 mL) andbrine (10 mL), dried (MgSO₄) and concentrated in vacuo to afford theproduct (145 mg) as a colorless foam which was used crude in thefollowing reaction. R_(f) (9:1:0.1, CH₂ Cl₂ :MeOH:NEt₃)=0.52; MS (ESI)m/z 777.4 (MH⁺).

Step 2

A mixture of the coupled product from Step 1 (147 mg, 0.189 mmol) and10% palladium on carbon (20 mg) in 1:1 methanol: acetic acid (6 mL) wasshaken under hydrogen at 50 psi. After 3 h the solids were removed byfiltration through celite and the filtrate concentrate in vacuo toafford the compound as a tan solid (111 mg) which was used crude in thenext step. R_(f) (9:1:0.1, methylene chloride:methanol:triethylamine)=0.26; ¹ HNMR (300 MHz, CD₃ OD) d, 7.36(t, 2H), 7.20(d, 1H), 6.8-7.01(m,9H), 3.03-3.28(m, 5H), 2.12(s, 3H), 1.34-1.91 (m, 24H); MS (ESI) m/z643.4 (MH⁺).

Step 3

Triethylamine (48 mL) was added to a solution of the above intermediate(111 mg, 0.172 mmol) in methylene chloride (2mL) followed by succinicanhydride (18.11 mg, 0.181 mmol) and the resultant solution stirred atroom temperature for 16 h. The solvents were removed in vacuo and theresidue purified by flash column chromatography (9:1:0.1, methylenechloride:methanol:acetic acid) as the eluant. The product was purifiedby preparative RP-HPLC (80-100% B in A, 30 min) to give 27.3 mg as afluffy pink lyophile. The lyophile was >98% pure as determined onanalytical RP-HPLC: tr=17.73 and 18.22 (80-100% B in A, 30 min). R_(f)(9:1:0.1, methylene chloride:methanol:acetic acid)=0.33; MS (ESI) m/z743.4 (MH⁺)

EXAMPLE 7N-[1S-[2-Adamantan-2-yl-oxy)carbonyl]-amino-3-(1H-indol-3-yl)-2S-methyl-propionylamino-methyl]-5-(3-o-tolyl-ureido)-pentyl]-succinamicacid

This was prepared analogously to Example 6. Hence Intermediate S (200mg, 0.504 mmol) and Intermediate G (200 mg, 0.504 mmol) gave the product(47.6 mg) as a white lyophile after purification by preparative RP-HPLC(60-90% B in A, 30 min). The lyophile was >98% pure as determined onanalytical RP-HPLC: tr=23.83 min (60-90% B in A, 30 mins); ¹ HNMR (300MHz, CDCl₃) d 8.42(b, 1H), 7.6(d, 1H), 7.0- 7.4(m, 11H), 6.a2(t, 1H),5.41(b, 1H), 4.82(b, 1H), 3.60(b, 1H), 3.2(b,2H), 2.62(b,2H), 2.38(b,2H), 2.22(s, 3H), 1.2-2.0(m, 24H); MS (ESI) m/z 743.4 (MH⁺).

EXAMPLE 8N-[1S-[2-Adamantan-2-yl-oxy)carbonyl]-amino-3-(1H-indol-3-yl)-2R-methyl-propionylamino-methyl]-5-(3-o-tolyl-ureido)-pentyl]-succinamicacid

This was prepared analogously to Example 6. Hence Intermediate S (200mg, 0.504 mmol) and Intermediate F (200 mg, 0.504 mmol) gave the product(35.2 mg) as a white lyophile after purification by preparative RP-HPLC(60-90% B in A, 30 min). The material was >98% pure, as determined onanalytical RP-HPLC: tr=22.64 min (60-90% B in A, 30 min); ¹ HNMR (300MHz, CDCl₃) d 8.50(b, 1H), 7.57(d, 1H), 6.85-7.4((m, 10H), 5.46(b, 1H),4.8(b, 1H), 3.8(b, 1H), 3.2(b, 2H), 2.6(b,2H), 2.38(b,2H), 2.23(s, 3H),1.4-2.0(m, 24H); MS (ESI) m/z 743.4 (MH⁺).

EXAMPLE 9N-[1S[2S-Adamantan-2-yl-oxy)carbonyl]-amino-3-(1H-indol-3-yl)-propionylamino-methyl]-5-(3-o-tolyl-ureido)-pentyl]-succinamic acid

Step 1

This was prepared in a similar manner to Example 6. Hence intermediate S(78.0 mg, 0.196 mmol) and intermediate J (75.0 mg, 0.196 mmol) gave theproduct (107.4 mg) as a tan solid which was used crude in the nextreaction. R_(f) (9:1:0.1, CH₂ Cl₂ :MeOH:NEt₃) =0.57; MS (ESI) m/z 763.4(MH⁺).

Step 2

This was prepared in a similar manner to step 2 of Example 6. Hence thematerial from step 1 (106.0 mg, 0.138 mmol) gave the product (97 mg) asa tan solid which was used crude in the next reaction; R_(f) (9:1:0.1,methylene chloride:methanol:triethylamine) =0.25.

Step 3

This was prepared in a similar manner to Example 6. Hence the materialfrom step 2 (117 mg, 0.169 mmol) gave the product (19.8 mg) as a whitelyophile. This was >98% pure on RP-HPLC: (80-100% B in A, 30 min); R_(f)(9:1:0.1, methylene chloride:methanol:triethylamine) =0.35; ¹ HNMR (300MHz, CDCl₃), d 8.35(b, 1H), 7.54(b, 1H), 7.10-7.35(m, 9H), 6.86(t, 1H),5.4(b, 1H), 4.82(b, 1H), 3.52(b, 1H), 3.22(b,2H), 2.59(b, 2H), 2.62(b,2H), 2.43(b, 2H), 2.22(b, 3H), 1.2-2.0(m, 22H); MS (ESI) m/z 792.2(MH+).

EXAMPLE 10N-[1S-[2R-Adamantan-2-yl-oxy)carbonyl]-amino-3-(1H-indol,3-yl)-propionylamino-methyl]-5-(3-o-tolyl-ureido)-pentyl]-succinamic acid

Step 1

This was prepared in a similar manner to Example 6. Hence, IntermediateS (78.0 mg, 0.196 mmol) and Intermediate H (75.0 mg, 0.196 mmol) gavethe product (138 mg) as a colorless foam which was used crude in thefollowing reaction; R_(f) (9:1:0.1; CH₂ CI₂ :MeOH:NEt₃) =0.56; MS (ESI)m/z 763.2 (MH⁺).

Step 2

This was prepared in a similar manner to step 2 of Example 6. Hence thematerial from step 1 (135 mg, 0.177 mmol) gave the product (89.9 mg) asa tan solid which was used crude in the next reaction; R_(f) (9:1:0.1,CH₂ CI₂ :MeOH:Et₃ N)=0.25; MS (ESI) m/z 629.4 (MH⁺).

Step 3

This was prepared in a similar manner to Example 6. Hence the materialfrom step 2 (88.0 mg, 0.127 mmol) gave the product (21.8 mg) as a whitelyophile, which was >98% pure on RP-HPLC: tr=5.32 min (80-100% B in A,30 min). R_(f) (9:1:0.1, methylene chloride:methanol:acetic acid)=0.35;¹ HNMR (300 MHz, CDCl₃), d 8.4(b, 1H), 7.59(d, 1H), 7.0-7.40 (m, 11H),6.83 (t, 1H), 5.4(b, 1H), 4.82(b, 1H), 3.6(b, 1H), 3.2(b, 2H), 2.62(b,2H), 2.4(b, 2H), 2.25(s, 3H), 1.2-2.0(m, 20H); MS (ESI) m/z 729.2((MH⁺).

EXAMPLE 11N-[1R-[2-Adamantan-2-yl-oxy)carbonyl-amino-3-(1H-indol-3-yl)-2RS-methyl-propionylamino-methyl]-4-(3-o-tolyl-ureido)-butyl]-succinamicacid

This was prepared in a analogous manner to Example 6. Hence IntermediateU (100 mg, 0.262 mmol) and Intermediate E (103 mg, 0.262 mmol) gave theproduct (103.0 mg) as a white lyophile after purification by RP-HPLC(60-90% B in A, 30 min); The lyophile was >98% pure as determined onanalytical RP-HPLC: tr=22.95 min (60-90% B in A, 30 min); MS (ESI) m/z729.4 (MH⁺).

EXAMPLE 12N-[1S-[2-Adamantan-2-yl-oxy)carbonyl-amino-3-(1H-indol-3-yl)-2RS-methyl-propionylamino-methyl]-4-(3-o-tolyl-ureido)-butyl]-succinamicacid

Step 1

This was prepared in a similar manner to Example 6. Hence Intermediate W(80.0 mg, 0.228 mmol) and Intermediate E (90.7 mg, 0.228 mmol) gave theproduct (142 mg) as a white powder which was used crude in the nextreaction. R_(f) (9:1:0.1; CH₂ Cl₂ :MeOH:NEt₃)=0.51; MS (ESI) m/z 729(MH⁺).

Step 2

The material from step 1 (140 mg, 0.192 mmol) was stirred in 4N hydrogenchloride in dioxane (2 mL) at room temperature for 2 h. Ether (10 mL)was added and the resultant solid isolated by filtration to afford theproduct (84 mg) as a white solid which was used crude for the nextreaction. R_(f) (9:1:0.1, methylene chloride:methanol:triethylamine)=0.28; MS (ESI) m/z 629.4 (MH⁺).

Step 3

This was prepared in a similar manner to Example 6. Hence the materialfrom step 2 (84.0 mg, 0.126 mmol) gave the crude product. The productwas purified by preparative RP-HPLC (75-100% B in A, 30 minutes) andisolated as a white lyophile. Analytical RP-HPLC showed an equal mixtureof diastereomers, tr=11.46 and 11.78 min (75-100% B in A, 30 min); R_(f)(9:1:0.1, methylene chloride:methanol:acetic acid)=0.32; MS (ESI) m/z729.2 (MH⁺);

EXAMPLE 13N-[1R-[2R-Adamantan-2-yl-oxy)carbonyl]-amino-3-(1H-indol-3-yl)propionylamino-methyl]-4-(3o-tolyl-ureido)-butyl]-succinamic acid

Step 1

This was prepared in a similar manner to Example 6. Hence Intermediate U(100 mg, 0.262 mmol) and Intermediate H (100 mg, 0.262 mmol) gave theproduct (180 mg) as a clear glass. This was used crude in the nextreaction. R_(f) (9:1:0.1; CH₂ Cl₂ :MeOH:NEt₃)=0.55; MS (ESI) m/z 749.4(MH⁺).

Step 2

This was prepared in a similar manner to step 2 from Example 6. Hencethe material from step 1 (180 mg, 0.240 mmol) gave the product (142 mg)as a tan solid which was used crude in next reaction. MS (ESI) m/z 615.1(MH⁺).

Step 3

This was prepared in a similar manner to Example 6. Hence the materialfrom step 2 (142 mg, 0.235 mmol) gave the product (50.5 mg) as a palepink lyophile after purification by preparative RP-HPLC (60-90% B in A,30 min). The material was >98% pure as determined on analytical RP-HPLC:tr =23.72 min (60-90% B in A, 30 min), ¹ HNMR (300 MHz, CDCl₃), d9.20(b, 1H), 7.61(b, 1H), 7.0-7.40(m, 9H), 6.92(t, 1H), 5.4(b, 1H),4.80(b, 1H), 3.49(b, 1H), 2.9-3.2(b, 4H), 2.61(b, 2H), 2.20(s,3H),1.2-20(m, 18H); MS (ESI) m/z 715.4 (MH⁺).

EXAMPLE 14N-[1R-[2S-Adamantan-2-yl-oxy)carbonyyl-amino-3-(1H-indol-3-yl)-propionylamino-methyl]-4-(3-o-tolyl-ureido)-butyl]-succinamic acid

Step 1

This was prepared in a similar manner to step 1 of Example 9. HenceIntermediate U (100 mg, 0.262 mmol) and Intermediate J (100 mg, 0.262mmol) gave the product as a colorless glass (161 mg) which was also usedcrude in following reaction. R_(f) (9:1:0.1; CH₂ Cl₂ :MeOH:NEt₃)=0.55;MS (ESI) m/z 749.4 (MH⁺).

Step 2

This was prepared in an analogous manner to Example 6. Hence thematerial from step 1 (161 mg, 0.215 mmol) gave the product (49.3 mg) asa white lyophile after purification by preparative RP-HPLC (60-90% B inA, 30 mins). The material was >98% pure as determined on analyticalRP-HPLC: tr=22.87 min (60-90%B in A, 30 mins), ¹ HNMR (300 MHz, CDCl₃) d8.78(b, 1H), 7.62(d, 1H), 7.1-7.45(m, 9H), 6.75(b, 1H), 5.69(b, 1H),4.80(s, 1H), 4.56(b, 1H), 3.1-3.6(water peak), 2.60(b, 2H), 2.25(s, 3H),1.05-1.95(m, 18H); MS (ESI) m/z 715.4 (MH⁺).

EXAMPLE 15N-[1S,[2S-Adamantan-2-yl-oxy)carbonyl]-amino-3-(1H-indol-3-yl)-propionylamino-methyl]-4-(3-o-tolyl-ureido)-butyl]-succinamic acid

This was prepared in a similar manner to Example 6. Hence Intermediate V(150 mg, 0.392 mmol) and Intermediate J (150 mg, 0.392 mmol) gave theproduct (46.0 mg) as a white lyophile after purification by preparativeRP-HPLC (60-90%B in A, 30 min). The material was >98% pure as determinedon analytical RP-HPLC: tr=21.96 min (60-90%B in A, 30 min). ¹ HNMR (300MHz, CDCl₃) d 9.38(b, 1H), 7.54(d, 1H), 6.9-7.4(m, 9H), 6.86(b, 1H),5.70(b, 1H), 4.4e(b, 1H), 3.60(b, 2H), 3.16(b, 2H), 2.6(b, 2H), 2.26(s,3H), 1.05-1.95(m, 18H); MS (ESI) m/z 715.4 (MH⁺).

EXAMPLE 16 N-[1S-[2R-Adamantan-2-yl-oxy)carbonyl]-amino-3-(1H-indol-3-yl)-propionylamino-methyl]-4-(3-o-tolyl-ureido)-butyl]-succinamic acid

This was prepared in a similar manner to Example 6. Hence Intermediate V(150 mg, 0.392 mmol) and Intermediate H (150 mg, 0.392 mmol) gave theproduct (134 mg) as a white lyophile after purification by preparativeRP-HPLC (60-90%B in A, 30 min). The material was >98% pure on analyticalRP-HPLC: tr=20.46 min (60-90%B in A, 30 min). ¹ HNMR (300 MHz, CDCl₃) d8.98(b, 1H), 7.70(m, 1H), 6.9-7.5(m, 9H), 6.90(b, 1H), 5.62(b, 1H),4.52(b, 1H), 3.60(b, 1H), 2.8-3.5(H₂ O peak), 2.50(b,2H), 2.22(s, 3H),1.02-1.95(m, 18H); MS (ESI) m/z 715.4 (MH⁺).

EXAMPLE 17N-[1R-[1-(Adamantan-2-yl-oxy)carbamoyl)-2-(1H-indol-3-yl)-1RS-methyl-ethyl-carbamoylmethyl]- 5-(3-o-tolyl-ureido)-pentyl]-succinamic acid

Step 1

To a solution of Intermediate N (0.169 g, 0.43 mmol), and Intermediate R(0.176 g, 0.43 mmol), N-hydroxybenzotriazole (0.058 g, 0.43 mmol) inN,N-dimethylformamide (5 mL) was added BOP reagent (0.19 g, 0.43 mmol).The reaction mixture was stirred overnight. The DMF was removed in vacuoand the residue was partitioned between ethyl acetate (20 mL) and 1Naqueous NaOH (20 mL). The organic layer was separated and washedsuccessively with water (20 mL), 10% aqueous citric acid (20 mL), andwater (5 mL), dried over sodium sulfate and evaporated in vacuo to anamorphous solid (0.273 g, 0.37 mmol) which was used without furtherpurification. R_(f) (250:25:1, methylene chloride methanol:aceticacid)=0.39; MS (FAB) m/z 727 (MH⁺)

Step 2

The product from the previous step (200 mg, 0.28 mmol) was dissolved in4N HCl in 1,4-dioxane (10 mL) and 0.2 mL anisole and stirred at 0° C.for 2 h. The 1,4-dioxane was evaporated and the residual oil wastriturated with ethyl acetate to afford the amine hydrochloride salt asan amorphous pink solid (157 mg, 0.25 mmol); MS (FAB) m/e 627 (MH⁺).

Step 3

The product from the previous step (155 mg, 0.25 mmol) was suspended inethyl acetate (10 mL) and triethylamine (0.048 mL, 0.25 mmol) andsuccinic anhydride (0.023 g, 0.25 mmol) was added. The mixture wassonicated 15 minutes, then allowed to stir at room temperatureovernight. The reaction mixture was diluted with ethyl acetate (20 mL),washed with 10% aqueous citric acid (20 mL), water (20 mL), dried (Na₂SO₄) and concentrated in vacuo to give an amorphous solid. Example 17was purified by preparative RP-HPLC: tr=24.0 min (60-90% B in A, 30min). On analytical RP-HPLC the diastereomers overlapped at tr=16.5 min(30-60%B in A); MS FAB m/e 727 (MH⁺).

EXAMPLE 18N-[1R-[2-[Benzloxy)carbonyl]-amino-3-(1H-indol-3-yl)-2RS-methyl-propionylamino-methyl]-5-(3-o-tolyl-ureido)-pentyl]-succinamic acid

Step 1

The product was obtained from the coupling of Intermediate K (0.352 g,0.0013 mol) and Intermediate O (0.49 g, 0.0013 mol) using EDC/HOBt aspreviously described. The crude material (0.917 g) was used in the nextstep without further purification. R_(f) (9:1, methylenechloride:methanol)=0.44; MS (FAB) m/e 699 (MH⁺).

Step 2

The product from the previous step was dissolved in 4N HCl dioxane (10mL) and anisole (0.2 mL) and stirred for two hours at 0° C. The dioxanewas evaporated in vacuo and the residue was triturated with ether togive the product as a white amorphous solid (723 mg). R_(f) (9:1,methylene chloride:methanol)=0.09; MS(FAB) m/e 599 (MH⁺).

Step 3

The product from the previous step (0.723 g, 1.20 mmol) was suspended inethyl acetate (50 mL) and triethylamine (0.167 mL) and succinicanhydride (0.114 g, 0.0012 mol) were added. The mixture was sonicatedfor 30 minutes, then allowed to stir overnight. The reaction mixture wasdiluted with ethyl acetate (50 mL) and washed with 10% aqueous citricacid and water. The organic layer was dried (Na₂ SO₄) and concentratedin vacuo to give 0.712 g of crude product. R_(f) (250:24:1, methylenechloride:methanol:acetic acid)=0.31. A portion of this material (147 mg)was purified by preparative RP-HPLC (50-80% B in A, 30 min). The productwas collected and lyophilized to an amorphous white solid. On analyticalRP-HPLC, the product diastereomers elute as a broad single peak: tr=19.8min (50-80% B in A, 30 min). MS (FAB) m/e 699 (MH⁺).

Intermediate Example i N-[1R-[2-Amino-3-(1H-indol-3-yl)-2RS-methyl-propionylamino-methyl]-5-(3-o-tolyl-ureido)-pentyl]-succinamic acid

To a portion of Example 18 (350 mg) dissolved in glacial acetic acid (10mL) and methanol (5 mL) was added 10% palladium on carbon (50 mg). Thismixture was shaken under a 50 psi H₂ atmosphere overnight, then filteredthrough a celite pad. The celite pad was washed with methanol (100 mL)and the filtrate was concentrated in vacuo. A portion of this materialwas purified by preparative RP-HPLC (40%-70%B in A, 30 min). The productwas collected and lyophilized to an amorphous white solid. A slightseparation of diastereomers could be observed on analyticalRP-HPLC:tr=7.8 min and 8.0 min (40-70%B in A, 30 min). R_(f) (250:25:1,methylene chloride: methanol:acetic acid)=0.02; MS (FAB) m/e 565 (MH⁺).

EXAMPLE 19 N-[1R-[2-Acetylamino-3-(1H-indol-3-yl)-2RS-methylpropionylamino-methyl]-5-(3-o-tolyl-ureido)-pentyl]-succinamic acid

A portion of the product of Intermediate Example i (0.05 g) wasdissolved in a solution of DMF (1 mL), pyridine (1 mL) and aceticanhydride (1 mL) and stirred for 15 minutes at room temperature. Thesolvent was concentrated in vacuo and the material purified bypreparative RP-HPLC (40-70%B in A, 30 min). The product was collectedand lyophilized to an amorphous white solid. No separation ofdiastereomers was observed on analytical RP-HPLC:tr=15 min (40-70%B inA, 30 min). MS (FAB) m/e 591 (MH⁺).

We claim:
 1. A compound of the following Formula (I): ##STR23## wherein:A is selected from--COOR¹, --CONHR¹, --CN, ---- tetrazolyl, --CONHSO₂R₃, --CH₂ OR¹, --CH₂ SR¹, --P--O(OR¹)₂ or --CO--NHOH; R¹ is H or C₁₋₆alkyl; R² is C₁₋₆ alkyl, CF₃, OH, or phenyl; R³ is OH, NH₂, or CH₃ ; R⁴is CN, CO₂ H or CF₃ ; B is selected from--COCH(NHR⁵)--(CH₂)_(n) --,--COCH₂ (CH₂)_(n) --, --CO--CH═CH--, --CO--phenylene, ortho, meta orpara, --CH₂ --(CH₂)_(n) -- or --CH(COOR⁶)--CH₂ --(CH₂)_(n) --; where nis 0, 1 or 2; R⁵ is H, C₁₋₆ alkyl, COR⁷ or CONHR⁸ ; R⁶ is H, C₁₋₆ alkyl,t-butyl, benzyl, methyl, ethyl or phenyl; R⁷ is H, C₁₋₆ alkyl, benzyl,C₁₋₆ alkoxy, or benzyloxy; R⁸ is H, C₁₋₆ alkyl, benzyl or phenyl; R⁹ isH or C₁₋₆ alkyl; R¹⁰ is H or C₁₋₆ alkyl; m is 1, 2, 3 or 4; G is --O--,--CH₂ --, NR¹¹, --CH═CH--; R¹¹ is H or C₁₋₆ alkyl; R¹² is selectedfromC₁₋₁₀ alkyl, C₂₋₆ alkenyl, C₃ -C₁₀ cycloalkyl, C₃ -C₁₀ carbocyclicarylalkyl or C₃ -C₁₀ carbocyclic aryl having 1 or 2 substituentsindependently selected from the group consisting ofhydroxyl, halogen--OSO₃ R¹³, nitro, cyano, amino, C₁₋₆ alkylamino, (C₁₋₆ alkyl)₂ amino,C₁₋₆ alkyl, halo C₁₋₆ alkyl C₁₋₆ alkoxy, C₂ -C₄ - alkanoyl, C₁₋₆ alkoxycarbonyl, and phenoxy; R¹³ is H or C₁₋₆ alkyl; p is 0, 1 or 2; D isselected from--NH--CO--, --CO--NR¹⁴ --, --(CH₂)_(r) --NR¹⁴ -- or --NR¹³--(CH₂)_(r) --; r is 0, 1 or 2; R¹⁴ is H or C₁₋₆ alkyl; R¹⁵ is H or C₁₋₆alkyl; q is 0 or 1; E is selected from--NH--CO--, --CO--NR¹⁷,--NH--CO--NR¹⁷ --, --O--CO--NR¹⁷, --SO₂ --NR¹⁷ --or --(CH₂)_(r) --NR₁₇--; R¹⁷ is H is C₁₋₁₀ alkyl; R¹⁸ is selected fromC₃ -C₁₀ carbocyclicarylalkyl, C₃ -C₁₀ carbocyclic aryl, C₁₋₁₀ alkyl, C₂₋₁₀ alkenyl or C₃-C₁₀ mono, bi- or tri-cycloalkyl with zero to four substituentsindependently selected form the group consisting of:C₁₋₁₀ alkyl,halogen, CN, --OR¹⁹, --SR¹⁹, --CO₂ R¹⁹, --CF₃, --NR¹⁹ R²⁰, --(CH₂)_(s)OR¹⁹ or --(CH₂)_(s) COOR¹⁹ ; s is an integer from 0 to 6; R¹⁹ is H orC₁₋₆ alkyl; and R²⁰ is H or C₁₋₆ alkyl; or a pharmaceutically acceptablebase-addition salt thereof.
 2. The compound of claim 1, whereinA isCOOR¹ ; B is --COCH² --(CH₂)_(n) ; R⁹ is H or C₁₋₆ alkyl; m is 3 or 4R¹⁰ is H or C₁₋₆ alkyl; G is NR¹¹ ; R¹² is C₃₋₁₀ carbocyclic aryl; p is1; D is --NHCO-- or --CONR¹⁴ ; R¹⁵ is H or C₁₋₆ alkyl; q is 1; E is--NHCO-- or --OCONR¹⁷ ; and R¹⁸ is C₃ -C₁₀ mono, bi or tri-cycloalkylwith 0 to 4 substituents.
 3. The compound of claim 1, wherein A isCOOR¹.
 4. The compound of claim 1; wherein B is --COCH₂ --(CH₂)_(n) andn=1 or
 2. 5. The compound of claim 1, wherein G is NR¹¹.
 6. The compoundof claim 1, wherein R¹² is aryl having 1 or 2 substituents.
 7. Thecompound of claim 1, wherein D is --NHCO-- or --CONR¹⁴.
 8. The compoundof claim 1, wherein E is --NHCO-- or --OCONR¹⁷.
 9. The compound of claim1, wherein R¹⁸ is C₃ -C₁₀ mono, bi or tri-cycloalkyl with 0 to 4substituents.
 10. The compound of claim 1, whereini) --B--A is --CO--CH₂CH₂ COOH or --COCH₂ (CH₂)_(n) NHS(O₂)CF₃ wherein n is 0 or 1; ii)--G--R¹² is --NH phenyl with Cl, OCH₃ or Ch₃ substitution at the orthoposition; and/or iii) R¹⁸ is adamantane, norbornane, 2-methylcyclohexylor tert-butyl.
 11. A CCK modulating composition comprising apharmaceutically-acceptable carrier and a therapeutically-effectiveamount of a compound according to claim
 1. 12. A method of mimicking orantagonizing the effects of CCK on CCK type-A receptors comprisingadministering to a mamalian host in need of such treatment atherapeutically-effective amount of a compound of claim 1.